sheep anti digoxigenin fab fragments antibody ap conjugated  (Roche)

 
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    Structured Review

    Roche sheep anti digoxigenin fab fragments antibody ap conjugated
    Patterns of Pax9 , Dkk1 and Dkk2 expression in E13.5 Pax9+/− and Pax9−/− palatal shelves suggest that Pax9 is upstream of Wnt signaling. (A,B) Whole-mount lacZ staining (blue) showing that Dkk1 expression is restricted to the posterior domain of palatal shelves in Dkk1Tg107−lacZ embryos at E13.5 and E14.5. Red dashed lines indicate the boundary of palate. (C,D) Whole-mount in situ hybridization with <t>digoxigenin-labeled</t> probes showing that Dkk2 transcripts (purple) are more visible at the medial edges of the palatal shelves in Pax9+/+ embryos at E13.5 and E14.5. (E,F) By contrast, Pax9 expression appears stronger and expands along the AP domain of palatal shelves at E13.5 and E14.5. Black dashed lines indicate the position of the sections in G-L. (G-L) In situ hybridizations on adjacent sections, presented in the coronal plane, confirm that Dkk1 (G,H) and Dkk2 (I,J) transcripts have inverse expression patterns with that of Pax9 (K,L) at E13.5 and E14.5. Pax9 expression is more intense along the buccal aspects of palatal mesenchyme (K,L), whereas Dkk2 transcripts localize on the lingual side (I,J). Arrows indicate the higher expression domain. (M-T) In the posterior regions of palatal mesenchyme, Dkk1 (M-P) and Dkk2 (Q-T) expression increased at E13.5 as a result of Pax9 deficiency. B, buccal; L, lingual; T, tongue. n =6 for whole-mount in situ hybridization, n =5 for section in situ hybridization.
    Sheep Anti Digoxigenin Fab Fragments Antibody Ap Conjugated, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 184 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Small-molecule Wnt agonists correct cleft palates in Pax9 mutant mice in utero"

    Article Title: Small-molecule Wnt agonists correct cleft palates in Pax9 mutant mice in utero

    Journal:

    doi: 10.1242/dev.157750

    Patterns of Pax9 , Dkk1 and Dkk2 expression in E13.5 Pax9+/− and Pax9−/− palatal shelves suggest that Pax9 is upstream of Wnt signaling. (A,B) Whole-mount lacZ staining (blue) showing that Dkk1 expression is restricted to the posterior domain of palatal shelves in Dkk1Tg107−lacZ embryos at E13.5 and E14.5. Red dashed lines indicate the boundary of palate. (C,D) Whole-mount in situ hybridization with digoxigenin-labeled probes showing that Dkk2 transcripts (purple) are more visible at the medial edges of the palatal shelves in Pax9+/+ embryos at E13.5 and E14.5. (E,F) By contrast, Pax9 expression appears stronger and expands along the AP domain of palatal shelves at E13.5 and E14.5. Black dashed lines indicate the position of the sections in G-L. (G-L) In situ hybridizations on adjacent sections, presented in the coronal plane, confirm that Dkk1 (G,H) and Dkk2 (I,J) transcripts have inverse expression patterns with that of Pax9 (K,L) at E13.5 and E14.5. Pax9 expression is more intense along the buccal aspects of palatal mesenchyme (K,L), whereas Dkk2 transcripts localize on the lingual side (I,J). Arrows indicate the higher expression domain. (M-T) In the posterior regions of palatal mesenchyme, Dkk1 (M-P) and Dkk2 (Q-T) expression increased at E13.5 as a result of Pax9 deficiency. B, buccal; L, lingual; T, tongue. n =6 for whole-mount in situ hybridization, n =5 for section in situ hybridization.
    Figure Legend Snippet: Patterns of Pax9 , Dkk1 and Dkk2 expression in E13.5 Pax9+/− and Pax9−/− palatal shelves suggest that Pax9 is upstream of Wnt signaling. (A,B) Whole-mount lacZ staining (blue) showing that Dkk1 expression is restricted to the posterior domain of palatal shelves in Dkk1Tg107−lacZ embryos at E13.5 and E14.5. Red dashed lines indicate the boundary of palate. (C,D) Whole-mount in situ hybridization with digoxigenin-labeled probes showing that Dkk2 transcripts (purple) are more visible at the medial edges of the palatal shelves in Pax9+/+ embryos at E13.5 and E14.5. (E,F) By contrast, Pax9 expression appears stronger and expands along the AP domain of palatal shelves at E13.5 and E14.5. Black dashed lines indicate the position of the sections in G-L. (G-L) In situ hybridizations on adjacent sections, presented in the coronal plane, confirm that Dkk1 (G,H) and Dkk2 (I,J) transcripts have inverse expression patterns with that of Pax9 (K,L) at E13.5 and E14.5. Pax9 expression is more intense along the buccal aspects of palatal mesenchyme (K,L), whereas Dkk2 transcripts localize on the lingual side (I,J). Arrows indicate the higher expression domain. (M-T) In the posterior regions of palatal mesenchyme, Dkk1 (M-P) and Dkk2 (Q-T) expression increased at E13.5 as a result of Pax9 deficiency. B, buccal; L, lingual; T, tongue. n =6 for whole-mount in situ hybridization, n =5 for section in situ hybridization.

    Techniques Used: Expressing, Staining, In Situ Hybridization, Labeling, In Situ

    2) Product Images from "Replication of avian, human and swine influenza viruses in porcine respiratory explants and association with sialic acid distribution"

    Article Title: Replication of avian, human and swine influenza viruses in porcine respiratory explants and association with sialic acid distribution

    Journal: Virology Journal

    doi: 10.1186/1743-422X-7-38

    Comparison of binding with digoxigenin-conjugated MAA in paraffin sections (A) and cryosections (B) of the porcine trachea . Only the cryosections showed clear positivity in the glands (black arrows) and the small blood vessels (blue arrows), while paraffin sections were completely negative.
    Figure Legend Snippet: Comparison of binding with digoxigenin-conjugated MAA in paraffin sections (A) and cryosections (B) of the porcine trachea . Only the cryosections showed clear positivity in the glands (black arrows) and the small blood vessels (blue arrows), while paraffin sections were completely negative.

    Techniques Used: Binding Assay

    Influence of the conjugation method of MAL-I and -II lectins on the staining intensities in duck small intestines . Biotinylated MAL-I (A) and MAL-II (a) both resulted in epithelial cell binding (black arrows), but MAL-II (a) was additionally staining the goblet cells (red arrow). For both lectins binding was abolished by sialidase treatment of the sections (B, b). Digoxigenin labelled MAL-I (C) and MAL-II (c) failed to bind to the same tissues.
    Figure Legend Snippet: Influence of the conjugation method of MAL-I and -II lectins on the staining intensities in duck small intestines . Biotinylated MAL-I (A) and MAL-II (a) both resulted in epithelial cell binding (black arrows), but MAL-II (a) was additionally staining the goblet cells (red arrow). For both lectins binding was abolished by sialidase treatment of the sections (B, b). Digoxigenin labelled MAL-I (C) and MAL-II (c) failed to bind to the same tissues.

    Techniques Used: Conjugation Assay, Staining, Binding Assay

    3) Product Images from "Differential effects of acute and chronic ethanol exposure on orexin expression in the perifornical lateral hypothalamus"

    Article Title: Differential effects of acute and chronic ethanol exposure on orexin expression in the perifornical lateral hypothalamus

    Journal:

    doi: 10.1111/j.1530-0277.2010.01161.x

    Photomicrographs illustrating the stimulatory effect of 0.75 g/kg ethanol on OX (a) mRNA expression and 2.5 g/kg ethanol on (b) peptide levels in the LH, as measured using digoxigenin-labeled ISH and immunofluorescence histochemistry as graphed in
    Figure Legend Snippet: Photomicrographs illustrating the stimulatory effect of 0.75 g/kg ethanol on OX (a) mRNA expression and 2.5 g/kg ethanol on (b) peptide levels in the LH, as measured using digoxigenin-labeled ISH and immunofluorescence histochemistry as graphed in

    Techniques Used: Expressing, Labeling, In Situ Hybridization, Immunofluorescence

    Effects of acute ethanol gavage on OX mRNA and peptide expression in the LH vs PF regions, as measured via (a) digoxigenin-labeled ISH and (b) immunofluorescence histochemistry (Experiment 4). The data (mean ±SEM) for the 3 groups (n=5/group),
    Figure Legend Snippet: Effects of acute ethanol gavage on OX mRNA and peptide expression in the LH vs PF regions, as measured via (a) digoxigenin-labeled ISH and (b) immunofluorescence histochemistry (Experiment 4). The data (mean ±SEM) for the 3 groups (n=5/group),

    Techniques Used: Expressing, Labeling, In Situ Hybridization, Immunofluorescence

    4) Product Images from "Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon "

    Article Title: Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon

    Journal:

    doi: 10.1128/IAI.71.2.985-990.2003

    Comparison of the patterns of binding of Stx and various lectins to mouse colon. Acetone-fixed frozen sections of the proximal ( < 4 cm below the cecum) (A, C, D, G, H, K, and L) or distal ( > 5 cm below the cecum) (B, E, F, I, J, M, and N) part of the colon were examined separately. The sections were stained with DIG-Stx1B plus HRP-anti-DIG (A, B, C, and E), biotin-GS-I-B4 (D and F), biotin-PNA (G and I), biotin-LEL (H and J), biotin-UEA-I (K and M), or biotin-AAL (L and N). The binding of biotin-conjugated lectins was revealed with HRP-avidin D. The distal part of the colon expressed binding sites for Stx1B (B and E). In contrast, the proximal part of the colon was not stained by Stx1B (A and C). Other lectins did not show a similar site-selective binding pattern. Arrowheads indicate the luminal surface. Arrows indicate the lamina propria (A, C, H, I, and J) or crypts of Lieberkühn (B, E, G, K, L, M, and N). Bars represent 25 μm (A and B [original magnification, ×360]) or 100 μm (C to N [original magnification, ×90]).
    Figure Legend Snippet: Comparison of the patterns of binding of Stx and various lectins to mouse colon. Acetone-fixed frozen sections of the proximal ( < 4 cm below the cecum) (A, C, D, G, H, K, and L) or distal ( > 5 cm below the cecum) (B, E, F, I, J, M, and N) part of the colon were examined separately. The sections were stained with DIG-Stx1B plus HRP-anti-DIG (A, B, C, and E), biotin-GS-I-B4 (D and F), biotin-PNA (G and I), biotin-LEL (H and J), biotin-UEA-I (K and M), or biotin-AAL (L and N). The binding of biotin-conjugated lectins was revealed with HRP-avidin D. The distal part of the colon expressed binding sites for Stx1B (B and E). In contrast, the proximal part of the colon was not stained by Stx1B (A and C). Other lectins did not show a similar site-selective binding pattern. Arrowheads indicate the luminal surface. Arrows indicate the lamina propria (A, C, H, I, and J) or crypts of Lieberkühn (B, E, G, K, L, M, and N). Bars represent 25 μm (A and B [original magnification, ×360]) or 100 μm (C to N [original magnification, ×90]).

    Techniques Used: Binding Assay, Staining, Avidin-Biotin Assay

    5) Product Images from "Fog1 is required for cardiac looping in zebrafish"

    Article Title: Fog1 is required for cardiac looping in zebrafish

    Journal:

    doi: 10.1016/j.ydbio.2005.10.040

    Expression of Fog genes during zebrafish development. Whole-mount in situ hybridization using digoxigenin-labeled antisense cRNA fog1 , fog2a , and fog2b probes on zebrafish embryos from 19 hpf to 72 hpf. An asterisk in the top left panel indicates intermediate
    Figure Legend Snippet: Expression of Fog genes during zebrafish development. Whole-mount in situ hybridization using digoxigenin-labeled antisense cRNA fog1 , fog2a , and fog2b probes on zebrafish embryos from 19 hpf to 72 hpf. An asterisk in the top left panel indicates intermediate

    Techniques Used: Expressing, In Situ Hybridization, Labeling

    6) Product Images from "Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation"

    Article Title: Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkt1343

    Cellular distribution of GFP different constructs RNA. ( A ) First column shows the GFP protein cellular localization of the different constructs where it can be seen that the signal is homogeneously distributed between nucleus and cytoplasm. ( B ) The second column shows the RNA cellular distribution of the constructs. Specific RNA signal is mainly cytoplasmic for the X7 and the X7-Δin7cDNA constructs, whilst it is present also in the nucleus for the X7-sup5′-3′ construct. Specific RNA detection was performed using anti-digoxigenin-rhodamin Fab fragments as described in the ‘Materials and Methods’ section. ( C ) Finally, the third column shows cells nuclei stained with TO-PRO-3. The panels on the right show the merge of these three signals. Scale bar 9 µm.
    Figure Legend Snippet: Cellular distribution of GFP different constructs RNA. ( A ) First column shows the GFP protein cellular localization of the different constructs where it can be seen that the signal is homogeneously distributed between nucleus and cytoplasm. ( B ) The second column shows the RNA cellular distribution of the constructs. Specific RNA signal is mainly cytoplasmic for the X7 and the X7-Δin7cDNA constructs, whilst it is present also in the nucleus for the X7-sup5′-3′ construct. Specific RNA detection was performed using anti-digoxigenin-rhodamin Fab fragments as described in the ‘Materials and Methods’ section. ( C ) Finally, the third column shows cells nuclei stained with TO-PRO-3. The panels on the right show the merge of these three signals. Scale bar 9 µm.

    Techniques Used: Construct, RNA Detection, Staining

    7) Product Images from "Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain"

    Article Title: Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

    Journal: Neural Development

    doi: 10.1186/1749-8104-6-10

    Inactivation of antibody-POD conjugate . Zebrafish embryos at 28 hpf were hybridized with dinitrophenyl-labeled shha and digoxigenin-labeled nkx6.1 RNA probes. (A, B) The expression patterns of shha (A) and nkx6.1 (B) as seen in single-color FISH experiments. In two-color experiments shha transcript was detected first using DyLight633-tyramide and nkx6.1 transcript was detected subsequently by FAM-tyramide. (C-H) Prior to the second round of detection, embryos were incubated for 10 minutes in PBS T (PBS plus 0.1% Tween-20) (C, D), PBS T containing 6% H 2 O 2 (E, F), or 100 mM glycine-HCl pH 2.0 (G, H). Single confocal sections of zebrafish brains are shown in the DyLight633-detection channel (Ch01) and in the FAM-detection channel (Ch02) from a lateral view and with anterior to the left. Images were recorded on a LSM510 microscope (Carl Zeiss) and false colored in ImageJ. Scale bar = 50 μm.
    Figure Legend Snippet: Inactivation of antibody-POD conjugate . Zebrafish embryos at 28 hpf were hybridized with dinitrophenyl-labeled shha and digoxigenin-labeled nkx6.1 RNA probes. (A, B) The expression patterns of shha (A) and nkx6.1 (B) as seen in single-color FISH experiments. In two-color experiments shha transcript was detected first using DyLight633-tyramide and nkx6.1 transcript was detected subsequently by FAM-tyramide. (C-H) Prior to the second round of detection, embryos were incubated for 10 minutes in PBS T (PBS plus 0.1% Tween-20) (C, D), PBS T containing 6% H 2 O 2 (E, F), or 100 mM glycine-HCl pH 2.0 (G, H). Single confocal sections of zebrafish brains are shown in the DyLight633-detection channel (Ch01) and in the FAM-detection channel (Ch02) from a lateral view and with anterior to the left. Images were recorded on a LSM510 microscope (Carl Zeiss) and false colored in ImageJ. Scale bar = 50 μm.

    Techniques Used: Labeling, Expressing, Fluorescence In Situ Hybridization, Incubation, Microscopy

    Positive effects of substituted phenol compounds on the TSA-POD reaction . Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for nkx6.1 are shown with anterior to the left. (A) Control (CTL); (B, C) vanillin added; (D, E) 4-iodophenol added. In all embryos shown, dextran sulfate was included in the hybridization and TSA-POD reaction. Above each panel signal-to-noise ratios (s/n) and compound concentrations are indicated. Black and white pictures were recorded with identical exposure times. Images were false-colored with help of the ImageJ software and no further adjustments or other image processing were performed. Scale bar = 100 μm.
    Figure Legend Snippet: Positive effects of substituted phenol compounds on the TSA-POD reaction . Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for nkx6.1 are shown with anterior to the left. (A) Control (CTL); (B, C) vanillin added; (D, E) 4-iodophenol added. In all embryos shown, dextran sulfate was included in the hybridization and TSA-POD reaction. Above each panel signal-to-noise ratios (s/n) and compound concentrations are indicated. Black and white pictures were recorded with identical exposure times. Images were false-colored with help of the ImageJ software and no further adjustments or other image processing were performed. Scale bar = 100 μm.

    Techniques Used: Labeling, CTL Assay, Hybridization, Software

    Bench-made fluorescent tyramides applied in zebrafish whole-mount FISH . Embryos were viewed on an Axioplan II microscope (Carl Zeiss). For excitation, a mercury burner (HBO 103 OSRAM) was used. Lateral views of zebrafish brains are shown with anterior to the left. Scale bar = 100 μm. ( A-C ) Zebrafish embryos at 28 hpf hybridized to a digoxigenin-labeled nkx6.1 antisense RNA probe. Images were captured with an Orca digital camera (Hamamatsu). As POD substrates, three different bench-made fluorogenic tyramides at a 1:250 dilution were used: (A) FAM-tyramide (exposure time, 100 ms; Chroma-filter 41001); (B) TAMRA-tyramide (exposure time, 4 ms; Chroma-filter 41002b); and (C) DyLight633-tyramide (exposure time, 120 ms; Chroma-filter 41008). ( D-F ) Zebrafish embryos at 24 hpf hybridized to a digoxigenin-labeled shha antisense RNA probe. Images were recorded with an Axiocam digital color camera (Carl Zeiss) using identical exposure times. Transcript distribution was visualized with (D) fluorescein-tyramide from Perkin Elmer (SAT701B001EA) at a 1:100 dilution, (E) bench-made FAM-tyramide at a 1:250 dilution, or (F) a 1:100 dilution.
    Figure Legend Snippet: Bench-made fluorescent tyramides applied in zebrafish whole-mount FISH . Embryos were viewed on an Axioplan II microscope (Carl Zeiss). For excitation, a mercury burner (HBO 103 OSRAM) was used. Lateral views of zebrafish brains are shown with anterior to the left. Scale bar = 100 μm. ( A-C ) Zebrafish embryos at 28 hpf hybridized to a digoxigenin-labeled nkx6.1 antisense RNA probe. Images were captured with an Orca digital camera (Hamamatsu). As POD substrates, three different bench-made fluorogenic tyramides at a 1:250 dilution were used: (A) FAM-tyramide (exposure time, 100 ms; Chroma-filter 41001); (B) TAMRA-tyramide (exposure time, 4 ms; Chroma-filter 41002b); and (C) DyLight633-tyramide (exposure time, 120 ms; Chroma-filter 41008). ( D-F ) Zebrafish embryos at 24 hpf hybridized to a digoxigenin-labeled shha antisense RNA probe. Images were recorded with an Axiocam digital color camera (Carl Zeiss) using identical exposure times. Transcript distribution was visualized with (D) fluorescein-tyramide from Perkin Elmer (SAT701B001EA) at a 1:100 dilution, (E) bench-made FAM-tyramide at a 1:250 dilution, or (F) a 1:100 dilution.

    Techniques Used: Fluorescence In Situ Hybridization, Microscopy, Labeling, Mass Spectrometry

    Optimization of hybridization and POD reaction by dextran sulfate . (A, C, E, G) Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for dbx1a are shown with anterior to the left. Above each panel signal-to-noise ratios (s/n) are indicated and whether dextran sulfate was added (+) or not (-) to the hybridization buffer (Hyb) and/or TSA reaction. (B, D, F, H) Embryos hybridized without probe were used as negative controls to assess background noise. Black-and-white pictures were recorded with identical exposure times using an Orca digital camera (Hamamatsu) on an Axioplan II microscope (Carl Zeiss). Images were false-colored with help of the ImageJ software and no further adjustments or other image processing was performed. Scale bar = 100 μm.
    Figure Legend Snippet: Optimization of hybridization and POD reaction by dextran sulfate . (A, C, E, G) Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for dbx1a are shown with anterior to the left. Above each panel signal-to-noise ratios (s/n) are indicated and whether dextran sulfate was added (+) or not (-) to the hybridization buffer (Hyb) and/or TSA reaction. (B, D, F, H) Embryos hybridized without probe were used as negative controls to assess background noise. Black-and-white pictures were recorded with identical exposure times using an Orca digital camera (Hamamatsu) on an Axioplan II microscope (Carl Zeiss). Images were false-colored with help of the ImageJ software and no further adjustments or other image processing was performed. Scale bar = 100 μm.

    Techniques Used: Hybridization, Labeling, Microscopy, Software

    8) Product Images from "Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems"

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems

    Journal: BMC Developmental Biology

    doi: 10.1186/1471-213X-11-43

    Control of bleed-through between Fast Blue and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Blue (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Blue (Ch01: detection of wavelengths greater than 650 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. No bleed-through was detected between the TSA-FAM and Fast Blue detection channels (B,C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.
    Figure Legend Snippet: Control of bleed-through between Fast Blue and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Blue (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Blue (Ch01: detection of wavelengths greater than 650 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. No bleed-through was detected between the TSA-FAM and Fast Blue detection channels (B,C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Techniques Used: Labeling, Fluorescence, Microscopy

    Fast Red and Fast Blue permit chromogenic and fluorescent transcript visualization . Lateral views of embryonic brains at 24 hpf hybridized to nkx6.1 digoxigenin and/or pax6a dinitrophenol antisense RNA probes as indicated on each panel. Transcript distributions were visualized by chromogenic (A,C,E) or fluorescent (B,D) detection of Fast Blue (C,D,E) and Fast Red (A,B,E) precipitates as indicated. Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Fluorescent signals were false-colored in ImageJ. Scale bar is 100 μm.
    Figure Legend Snippet: Fast Red and Fast Blue permit chromogenic and fluorescent transcript visualization . Lateral views of embryonic brains at 24 hpf hybridized to nkx6.1 digoxigenin and/or pax6a dinitrophenol antisense RNA probes as indicated on each panel. Transcript distributions were visualized by chromogenic (A,C,E) or fluorescent (B,D) detection of Fast Blue (C,D,E) and Fast Red (A,B,E) precipitates as indicated. Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Fluorescent signals were false-colored in ImageJ. Scale bar is 100 μm.

    Techniques Used: Microscopy

    Control of bleed-through between Fast Red and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Red (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Red (Ch01: detection of wavelengths greater than 560 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. A significant bleed-through of TSA-FAM nkx6.1 signal was detected in the Fast Red detection channel (C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.
    Figure Legend Snippet: Control of bleed-through between Fast Red and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Red (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Red (Ch01: detection of wavelengths greater than 560 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. A significant bleed-through of TSA-FAM nkx6.1 signal was detected in the Fast Red detection channel (C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Techniques Used: Labeling, Fluorescence, Microscopy

    Effects of dextran sulfate in standard WISH . 24-hpf embryos were hybridized to a sim1a digoxigenin-labeled RNA probe, which was visualized by AP-BCIP/NBT chromogenic detection under identical conditions and staining times. Lateral views of embryonic brains (A,B) and trunks (C,D) are shown with anterior to the left. In (B,D) but not in (A,C) 5% dextran sulfate was included in the hybridization buffer. Addition of dextran sulfate resulted in increased signal sensitivity. Arrowheads in (A,B) indicate sim1a -positive neuronal clusters identified in dextran sulfate treated brains (B) that were hardly detected in untreated embryos (A). Arrowheads in (C,D) mark the pronephric primordium strongly visualized in dextrane sulfate treated embryos (D) but not in untreated specimens (C). Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Scale bar is 100 μm.
    Figure Legend Snippet: Effects of dextran sulfate in standard WISH . 24-hpf embryos were hybridized to a sim1a digoxigenin-labeled RNA probe, which was visualized by AP-BCIP/NBT chromogenic detection under identical conditions and staining times. Lateral views of embryonic brains (A,B) and trunks (C,D) are shown with anterior to the left. In (B,D) but not in (A,C) 5% dextran sulfate was included in the hybridization buffer. Addition of dextran sulfate resulted in increased signal sensitivity. Arrowheads in (A,B) indicate sim1a -positive neuronal clusters identified in dextran sulfate treated brains (B) that were hardly detected in untreated embryos (A). Arrowheads in (C,D) mark the pronephric primordium strongly visualized in dextrane sulfate treated embryos (D) but not in untreated specimens (C). Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Scale bar is 100 μm.

    Techniques Used: Labeling, Staining, Hybridization, Microscopy

    9) Product Images from "TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis"

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis

    Journal:

    doi: 10.4161/spmg.1.1.15263

    Hgs, Zfyve9, Net25 and Smurf1 mRNAs and MAN1 and SMURF2 proteins are expressed in immature and adult testes. (A) Northern blot analysis of Hgs, Zfyve9, Net25 and Smurf1 transcripts in total RNA isolated from immature and adult mouse testes, detected using digoxigenin-labeled antisense riboprobes and indicated by black arrowheads. Transcript sizes were determined using an RNA ladder (left). Ethidium bromide stained gel (lower part) indicates the amount of RNA loaded per lane. (B) Western blot analysis of immature and adult mouse testis lysates detected bands of the expected sizes for MAN1 and SMURF2 proteins in whole testis lysates at the indicated ages. Asterisk indicates non-specific binding of secondary antibody used for MAN1 detection, as determined by blotting adult testis lysate in the absence of primary antibody (control, [C]). SMURF2 protein was detected at the expected size in 4 dpp mouse testis lysates and lysates prepared from whole 12.5 dpc fetus. Additional bands detected in adult testis lysates at 44, 72 and 130 kDa suggest the existence of different SMURF2 isoforms. No band was detected in adult testis lysates in the absence of anti-SMURF2 primary antibody [control, (C)]. Detection of α-TUBULIN (lower part) indicates amount of protein present in each lane.
    Figure Legend Snippet: Hgs, Zfyve9, Net25 and Smurf1 mRNAs and MAN1 and SMURF2 proteins are expressed in immature and adult testes. (A) Northern blot analysis of Hgs, Zfyve9, Net25 and Smurf1 transcripts in total RNA isolated from immature and adult mouse testes, detected using digoxigenin-labeled antisense riboprobes and indicated by black arrowheads. Transcript sizes were determined using an RNA ladder (left). Ethidium bromide stained gel (lower part) indicates the amount of RNA loaded per lane. (B) Western blot analysis of immature and adult mouse testis lysates detected bands of the expected sizes for MAN1 and SMURF2 proteins in whole testis lysates at the indicated ages. Asterisk indicates non-specific binding of secondary antibody used for MAN1 detection, as determined by blotting adult testis lysate in the absence of primary antibody (control, [C]). SMURF2 protein was detected at the expected size in 4 dpp mouse testis lysates and lysates prepared from whole 12.5 dpc fetus. Additional bands detected in adult testis lysates at 44, 72 and 130 kDa suggest the existence of different SMURF2 isoforms. No band was detected in adult testis lysates in the absence of anti-SMURF2 primary antibody [control, (C)]. Detection of α-TUBULIN (lower part) indicates amount of protein present in each lane.

    Techniques Used: Northern Blot, Isolation, Labeling, Staining, Western Blot, Binding Assay

    10) Product Images from "Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems"

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems

    Journal: BMC Developmental Biology

    doi: 10.1186/1471-213X-11-43

    Control of bleed-through between Fast Blue and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Blue (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Blue (Ch01: detection of wavelengths greater than 650 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. No bleed-through was detected between the TSA-FAM and Fast Blue detection channels (B,C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.
    Figure Legend Snippet: Control of bleed-through between Fast Blue and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Blue (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Blue (Ch01: detection of wavelengths greater than 650 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. No bleed-through was detected between the TSA-FAM and Fast Blue detection channels (B,C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Techniques Used: Labeling, Fluorescence, Microscopy

    Fast Red and Fast Blue permit chromogenic and fluorescent transcript visualization . Lateral views of embryonic brains at 24 hpf hybridized to nkx6.1 digoxigenin and/or pax6a dinitrophenol antisense RNA probes as indicated on each panel. Transcript distributions were visualized by chromogenic (A,C,E) or fluorescent (B,D) detection of Fast Blue (C,D,E) and Fast Red (A,B,E) precipitates as indicated. Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Fluorescent signals were false-colored in ImageJ. Scale bar is 100 μm.
    Figure Legend Snippet: Fast Red and Fast Blue permit chromogenic and fluorescent transcript visualization . Lateral views of embryonic brains at 24 hpf hybridized to nkx6.1 digoxigenin and/or pax6a dinitrophenol antisense RNA probes as indicated on each panel. Transcript distributions were visualized by chromogenic (A,C,E) or fluorescent (B,D) detection of Fast Blue (C,D,E) and Fast Red (A,B,E) precipitates as indicated. Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Fluorescent signals were false-colored in ImageJ. Scale bar is 100 μm.

    Techniques Used: Microscopy

    Control of bleed-through between Fast Red and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Red (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Red (Ch01: detection of wavelengths greater than 560 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. A significant bleed-through of TSA-FAM nkx6.1 signal was detected in the Fast Red detection channel (C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.
    Figure Legend Snippet: Control of bleed-through between Fast Red and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Red (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Red (Ch01: detection of wavelengths greater than 560 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. A significant bleed-through of TSA-FAM nkx6.1 signal was detected in the Fast Red detection channel (C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Techniques Used: Labeling, Fluorescence, Microscopy

    Effects of dextran sulfate in standard WISH . 24-hpf embryos were hybridized to a sim1a digoxigenin-labeled RNA probe, which was visualized by AP-BCIP/NBT chromogenic detection under identical conditions and staining times. Lateral views of embryonic brains (A,B) and trunks (C,D) are shown with anterior to the left. In (B,D) but not in (A,C) 5% dextran sulfate was included in the hybridization buffer. Addition of dextran sulfate resulted in increased signal sensitivity. Arrowheads in (A,B) indicate sim1a -positive neuronal clusters identified in dextran sulfate treated brains (B) that were hardly detected in untreated embryos (A). Arrowheads in (C,D) mark the pronephric primordium strongly visualized in dextrane sulfate treated embryos (D) but not in untreated specimens (C). Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Scale bar is 100 μm.
    Figure Legend Snippet: Effects of dextran sulfate in standard WISH . 24-hpf embryos were hybridized to a sim1a digoxigenin-labeled RNA probe, which was visualized by AP-BCIP/NBT chromogenic detection under identical conditions and staining times. Lateral views of embryonic brains (A,B) and trunks (C,D) are shown with anterior to the left. In (B,D) but not in (A,C) 5% dextran sulfate was included in the hybridization buffer. Addition of dextran sulfate resulted in increased signal sensitivity. Arrowheads in (A,B) indicate sim1a -positive neuronal clusters identified in dextran sulfate treated brains (B) that were hardly detected in untreated embryos (A). Arrowheads in (C,D) mark the pronephric primordium strongly visualized in dextrane sulfate treated embryos (D) but not in untreated specimens (C). Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Scale bar is 100 μm.

    Techniques Used: Labeling, Staining, Hybridization, Microscopy

    11) Product Images from "Active nuclear transcriptome analysis reveals inflammasome-dependent mechanism for early neutrophil response to Mycobacterium marinum"

    Article Title: Active nuclear transcriptome analysis reveals inflammasome-dependent mechanism for early neutrophil response to Mycobacterium marinum

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-06099-x

    In situ hybridisation reveals spatial analysis of M . marinum -induced upregulated genes. Embryos were injected at the 32-512-cell stage with M . marinum . Expression patterns and gene transcript levels were analysed by in situ hybridisation using digoxigenin-labelled RNA probes, in uninjected controls ( A – Li ) and M . marinum -injected embryos ( A – Lii ). Control expression patterns are indicated by the black arrows with black arrowheads used to indicate altered expression patterns for fos1a ( A ), junba ( B ), il81a ( C ), cepbp ( D ), junbb ( E ), hbegfa ( F ), egr4 ( G ), egr3 ( H ), ier2 ( I ), wu:fb15h11 ( J ), caspase b ( K ) and tnfb ( L ).
    Figure Legend Snippet: In situ hybridisation reveals spatial analysis of M . marinum -induced upregulated genes. Embryos were injected at the 32-512-cell stage with M . marinum . Expression patterns and gene transcript levels were analysed by in situ hybridisation using digoxigenin-labelled RNA probes, in uninjected controls ( A – Li ) and M . marinum -injected embryos ( A – Lii ). Control expression patterns are indicated by the black arrows with black arrowheads used to indicate altered expression patterns for fos1a ( A ), junba ( B ), il81a ( C ), cepbp ( D ), junbb ( E ), hbegfa ( F ), egr4 ( G ), egr3 ( H ), ier2 ( I ), wu:fb15h11 ( J ), caspase b ( K ) and tnfb ( L ).

    Techniques Used: In Situ, Hybridization, Injection, Expressing

    12) Product Images from "PRENATAL ETHANOL EXPOSURE STIMULATES NEUROGENESIS IN HYPOTHALAMIC AND LIMBIC PEPTIDE SYSTEMS: POSSIBLE MECHANISM FOR OFFSPRING ETHANOL OVERCONSUMPTION"

    Article Title: PRENATAL ETHANOL EXPOSURE STIMULATES NEUROGENESIS IN HYPOTHALAMIC AND LIMBIC PEPTIDE SYSTEMS: POSSIBLE MECHANISM FOR OFFSPRING ETHANOL OVERCONSUMPTION

    Journal:

    doi: 10.1016/j.neuroscience.2012.05.066

    Prenatal ethanol increases the density of peptide-expressing neurons in P0 offspring prenatally exposed to 1 g/kg or 3 g/kg ethanol relative to control ( n = 6–8/group), as assessed by in situ hybridization histochemistry with digoxigenin-labeled
    Figure Legend Snippet: Prenatal ethanol increases the density of peptide-expressing neurons in P0 offspring prenatally exposed to 1 g/kg or 3 g/kg ethanol relative to control ( n = 6–8/group), as assessed by in situ hybridization histochemistry with digoxigenin-labeled

    Techniques Used: Expressing, In Situ Hybridization, Labeling

    13) Product Images from "TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis"

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis

    Journal:

    doi: 10.4161/spmg.1.1.15263

    Hgs, Zfyve9, Net25 and Smurf1 mRNAs and MAN1 and SMURF2 proteins are expressed in immature and adult testes. (A) Northern blot analysis of Hgs, Zfyve9, Net25 and Smurf1 transcripts in total RNA isolated from immature and adult mouse testes, detected using digoxigenin-labeled antisense riboprobes and indicated by black arrowheads. Transcript sizes were determined using an RNA ladder (left). Ethidium bromide stained gel (lower part) indicates the amount of RNA loaded per lane. (B) Western blot analysis of immature and adult mouse testis lysates detected bands of the expected sizes for MAN1 and SMURF2 proteins in whole testis lysates at the indicated ages. Asterisk indicates non-specific binding of secondary antibody used for MAN1 detection, as determined by blotting adult testis lysate in the absence of primary antibody (control, [C]). SMURF2 protein was detected at the expected size in 4 dpp mouse testis lysates and lysates prepared from whole 12.5 dpc fetus. Additional bands detected in adult testis lysates at 44, 72 and 130 kDa suggest the existence of different SMURF2 isoforms. No band was detected in adult testis lysates in the absence of anti-SMURF2 primary antibody [control, (C)]. Detection of α-TUBULIN (lower part) indicates amount of protein present in each lane.
    Figure Legend Snippet: Hgs, Zfyve9, Net25 and Smurf1 mRNAs and MAN1 and SMURF2 proteins are expressed in immature and adult testes. (A) Northern blot analysis of Hgs, Zfyve9, Net25 and Smurf1 transcripts in total RNA isolated from immature and adult mouse testes, detected using digoxigenin-labeled antisense riboprobes and indicated by black arrowheads. Transcript sizes were determined using an RNA ladder (left). Ethidium bromide stained gel (lower part) indicates the amount of RNA loaded per lane. (B) Western blot analysis of immature and adult mouse testis lysates detected bands of the expected sizes for MAN1 and SMURF2 proteins in whole testis lysates at the indicated ages. Asterisk indicates non-specific binding of secondary antibody used for MAN1 detection, as determined by blotting adult testis lysate in the absence of primary antibody (control, [C]). SMURF2 protein was detected at the expected size in 4 dpp mouse testis lysates and lysates prepared from whole 12.5 dpc fetus. Additional bands detected in adult testis lysates at 44, 72 and 130 kDa suggest the existence of different SMURF2 isoforms. No band was detected in adult testis lysates in the absence of anti-SMURF2 primary antibody [control, (C)]. Detection of α-TUBULIN (lower part) indicates amount of protein present in each lane.

    Techniques Used: Northern Blot, Isolation, Labeling, Staining, Western Blot, Binding Assay

    14) Product Images from "Glutamatergic input to the lateral hypothalamus stimulates ethanol intake: Role of orexin and melanin-concentrating hormone"

    Article Title: Glutamatergic input to the lateral hypothalamus stimulates ethanol intake: Role of orexin and melanin-concentrating hormone

    Journal:

    doi: 10.1111/j.1530-0277.2012.01854.x

    (A) The stimulatory effect of glutamate on OX expression was confirmed using in situ hybridization with a digoxigenin-labeled probe (n = 5–6/group). In the LH, NMDA significantly increased the expression of OX in both PF and LH region, while AMPA
    Figure Legend Snippet: (A) The stimulatory effect of glutamate on OX expression was confirmed using in situ hybridization with a digoxigenin-labeled probe (n = 5–6/group). In the LH, NMDA significantly increased the expression of OX in both PF and LH region, while AMPA

    Techniques Used: Expressing, In Situ Hybridization, Labeling

    15) Product Images from "Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome"

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome

    Journal:

    doi: 10.1016/j.ijdevneu.2012.05.005

    Representative images from a female mouse at weaning age to illustrate digoxigenin-based Snord116 in situ hybridization (ISH) signal in multiple brain regions. Panels A (caudal diencephalon) and B (rostral mesencephalon) illustrate coronal sections hundreds
    Figure Legend Snippet: Representative images from a female mouse at weaning age to illustrate digoxigenin-based Snord116 in situ hybridization (ISH) signal in multiple brain regions. Panels A (caudal diencephalon) and B (rostral mesencephalon) illustrate coronal sections hundreds

    Techniques Used: In Situ Hybridization

    Representative images from a female mouse at weaning age show digoxigenin-based Snord116 in situ hybridization (ISH) signals in the hypothalamus. The ISH signal was detected in the paraventricular nucleus (PVN; panel A), ventromedial nucleus of the hypothalamus
    Figure Legend Snippet: Representative images from a female mouse at weaning age show digoxigenin-based Snord116 in situ hybridization (ISH) signals in the hypothalamus. The ISH signal was detected in the paraventricular nucleus (PVN; panel A), ventromedial nucleus of the hypothalamus

    Techniques Used: In Situ Hybridization

    Representative digoxigenin-based in situ hybridization (ISH) images of Snord116 from the rostral to caudal hypothalamus in a female mouse at weaning age. The images from panels A to L are hypothalamic sections taken from rostral to caudal. The Snord116
    Figure Legend Snippet: Representative digoxigenin-based in situ hybridization (ISH) images of Snord116 from the rostral to caudal hypothalamus in a female mouse at weaning age. The images from panels A to L are hypothalamic sections taken from rostral to caudal. The Snord116

    Techniques Used: In Situ Hybridization

    Digital images show digoxigenin-based in situ hybridization (ISH) illustrating the expression of Snord116 in the brain including hypothalamus and surrounding area in neonatal (P0) mice. Panels A and B show the representative images of Snord116 in mice
    Figure Legend Snippet: Digital images show digoxigenin-based in situ hybridization (ISH) illustrating the expression of Snord116 in the brain including hypothalamus and surrounding area in neonatal (P0) mice. Panels A and B show the representative images of Snord116 in mice

    Techniques Used: In Situ Hybridization, Expressing, Mouse Assay

    16) Product Images from ""

    Article Title:

    Journal:

    doi: 10.1074/jbc.M109.014506

    Expression of SLIP-GC in germinal center B cells. In situ hybridization using serially cut sections of the spleen of an immunized C57BL6 mouse is shown. Some sections were stained with PNA for GC B cells, and others were treated with digoxigenin-labeled antisense probe against SLIP-GC. Sense probe was used as a negative control. The numbered follicles with the PNA stain in the overview picture of the spleen are zoomed in and shown side by side with accompanying antisense and sense probes treated sections. The dark blue spots in the center of follicles 1 and 2 with the antisense probe depict cells that are positive for SLIP-GC expression, and these overlap with PNA positive ( dark red ) B cells. Follicle 3 lacks a signal with the antisense probe as well as PNA-positive cells because this is a follicle lacking a GC.
    Figure Legend Snippet: Expression of SLIP-GC in germinal center B cells. In situ hybridization using serially cut sections of the spleen of an immunized C57BL6 mouse is shown. Some sections were stained with PNA for GC B cells, and others were treated with digoxigenin-labeled antisense probe against SLIP-GC. Sense probe was used as a negative control. The numbered follicles with the PNA stain in the overview picture of the spleen are zoomed in and shown side by side with accompanying antisense and sense probes treated sections. The dark blue spots in the center of follicles 1 and 2 with the antisense probe depict cells that are positive for SLIP-GC expression, and these overlap with PNA positive ( dark red ) B cells. Follicle 3 lacks a signal with the antisense probe as well as PNA-positive cells because this is a follicle lacking a GC.

    Techniques Used: Expressing, Gas Chromatography, In Situ Hybridization, Staining, Labeling, Negative Control

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    Article Snippet: Embryos were washed for 15 min at 65 °C twice with 2X SSC, 0.1% Tween-20, twice with 0.2× SSC, 0.1% Tween-20, and at room temperature three times with PBT-Tween. .. Embryos were incubated with preabsorbed anti-Digoxigenin-alcaline phosphatase antibody (Roche, 11093274910) at 1:2000 for 1.5 h at room temperature on a wheel. .. They were washed three times for 20 min in PBT-Tween and once for 20 min in AP buffer (100 mM Tris 2 M pH 9.5, 50 mM MgCl2 , 100 mM NaCl, 0.1% Tween-20).

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: Briefly, for in situ hybridization, formalin-fixed and paraffin-embedded samples were deparaffinized, rehydrated, treated with HCl, digested in proteinase K solution, refixed with formaldehyde, treated in acetic anhydride solution, and hybridized with a digoxigenin labelled RNA probe specific for Ascl2 . .. After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. For immunofluorescent detection of Sox9, sections were treated as follows, at room temperature unless stated otherwise.

    Expressing:

    Article Title: Replication of avian, human and swine influenza viruses in porcine respiratory explants and association with sialic acid distribution
    Article Snippet: Paragraph title: -Expression of α2-6 linked Sia- ... Subsequently, the slides were incubated with 1:200 sheep alkaline phosphatase conjugated anti-digoxigenin Fab fragments (Roche) and developed with New Fuchsin (Dako).

    Article Title: PRENATAL ETHANOL EXPOSURE STIMULATES NEUROGENESIS IN HYPOTHALAMIC AND LIMBIC PEPTIDE SYSTEMS: POSSIBLE MECHANISM FOR OFFSPRING ETHANOL OVERCONSUMPTION
    Article Snippet: For better visualization, in situ hydridization histochemistry with a digoxigenin-labeled probe (digoxigenin-labeled in situ hybridization; DIG-ISH) was performed to reveal the anatomical distribution at birth (P0) of changes in peptide expression in the control, 1 g/kg ethanol and 3 g/kg dose ethanol groups ( n = 6–8/group) as described ( , ). .. AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000; Roche Diagnostics, Indianapolis, IN, USA) and NBT/BCIP (Roche Diagnostics) were used to visualize the signal.

    Article Title: Glutamatergic input to the lateral hypothalamus stimulates ethanol intake: Role of orexin and melanin-concentrating hormone
    Article Snippet: In Experiment 3, the mRNA expression of OX and MCH was also measured with digoxigenin (DIG)-labeled antisense RNA probes and 30-μm free-floating cryostat sections were used for ISH histochemistry as described previously ( ). .. AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000, Roche, Nutley, NJ) and NBT/BCIP (Roche, Nutley, NJ) were used to visualize the signal.

    Hybridization:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome
    Article Snippet: Before hybridization, sections were incubated in pre-hybridization solution (50% formamide in saline-sodium citrate (SSC; Sigma, S6639) buffer containing 8.8 units/ml heparin (Sigma, H9399), 0.05mg/ml yeast tRNA (Sigma, R5636), and 0.024mg/ml dextran sulfate (Sigma, D8906)) for 1h at 60°C. .. Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910). .. In situ hybridization was used to localize Hgs, Zfyve9, Smurf1 and Net25 transcripts in mouse testis sections.

    Article Title:
    Article Snippet: RNA probes were generated from the mouse IMAGE cDNA clone EMM1002-3872135 (Open Biosystems, Huntsville, AL), added to the hybridization buffer, hybridized to tissue sections overnight at 55 °C in a humidity chamber, and washed with 5×, 2×, and 0.2× SSC (1× SSC = 0.15 m NaCl and 0.015 m sodium citrate) buffer. .. The hybridized probe was detected after overnight incubation at 4 °C using an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche Applied Science catalog no. 11093274910).

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Hybridization was performed with 100–400 ng probe per slide at 50–60°C with stringency washes to 0.1× SSC at the hybridization temperature. .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042).

    Article Title: Active nuclear transcriptome analysis reveals inflammasome-dependent mechanism for early neutrophil response to Mycobacterium marinum
    Article Snippet: Embryos were equilibrated with hybridization buffer (50% Formamide, 1.3× SSC pH 5.0, 5 mM EDTA pH 8.0, 200 μg/mL Baker’s yeast tRNA, 0.2% Tween-20, 0.5% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 100 μg/mL Heparin) and incubated overnight with DIG-labeled probes at 68 °C). .. Probes were detected with an anti-DIG-alkaline phosphatase antibody (Roche, cat # 11093274910) and the signal was developed using BM Purple AP (Roche, cat # 11442074001).

    Article Title: Fog1 is required for cardiac looping in zebrafish
    Article Snippet: Embryos were then pre-hybridized in Hyb mix (5 × SSC, 65% formamide, 50 μg/ml heparin, 0.1% Tween-20, 0.5 mg/ml tRNA) for 3 h at 70°C followed by hybridization with the digoxigenin-labeled riboprobe for 16 h at 70°C. .. An alkaline phosphatase conjugated sheep anti-digoxigenin Fab fragment (Roche) was added (1:5000 dilution) and incubated for 16 h at 4°C.

    Article Title: Silencing microRNA-134 produces neuroprotective and prolonged seizure-suppressive effects
    Article Snippet: Paragraph title: In situ hybridization ... Then sections were rinsed in B1 buffer (150 mM NaCl, 100 mM Maleic acid and 0.4% IGEPAL pH:7.5) for 1 h at room temperature and B2 buffer (2% blocking reagent and 10% goat serum in B1) for 30 min. Polyclonal antibodies against digoxigenin (1:1000, Cat. 11093274910, Roche) were incubated in B2 overnight at 4 °C.

    TUNEL Assay:

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. For immunohistochemical detection of Ki67, sections were deparaffinized, rehydrated and subjected to antigen unmasking in the same fashion as for Sox9 staining, and were then treated with 2.25% hydrogen peroxide for 15 min, washed in water and PBST for 5 min each, blocked with Avidin D reagent (Vector Laboratories SP-2001), washed in PBST, blocked with 4% BSA in PBST for 30 min, incubated with rabbit anti-Ki67 antibody (Abcam Ab16667 at 1:100), washed three times for 5 min in PBST, incubated with biotinylated anti-rabbit antibody (Vector Laboratories BA-1000 at 7.5 μg ml−1 ) for 30 min, washed in PBST 3–5 min, stained with the Vector Laboratories ABC and DAB reagents (PK-6200 and SK-4100), counterstained with Gill #2 haematoxylin (Thermo Fisher Scientific), dehydrated in an ethanol series (70–100%), cleared in xylene, and mounted in Cytoseal XYL (Thermo Fisher Scientific 8312-4).

    Immunohistochemistry:

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. Sections were deparaffinized by heating to 60 °C for 15 min, washed 3 × 5 min in xylene, rehydrated in a series of ethanol solutions (100%, 95%, 90% and 70% ethanol), washed in water and in 0.1% Tween-20 for 1 min each, steamed in antigen unmasking solution (Vector Laboratories H-3300) for 15 min, washed in phosphate-buffered saline containing 0.2% Tween-20 (PBST) for 5 min, blocked in 4% BSA in PBST for 30 min, incubated with rabbit anti-Sox9 antibodies (Millipore AB5535 at 1.7 μg ml−1 ) overnight at 4° in 1% BSA in PBST, washed three times for 5 min in PBST, incubated with Alexa 488-conjugated goat anti-rabbit antibody (Invitrogen A-11034 at 2.5 μg ml−1 ) for 2 hours, washed three times for five minutes in PBST including 0.2 μg ml−1 4,6-diamidino-2-phenylindole (DAPI) in the penultimate wash, and mounted in ProLong Gold antifade reagent (Thermo Fisher Scientific).

    Southern Blot:

    Article Title: Inhibition of non-homologous end joining increases the efficiency of CRISPR/Cas9-mediated precise [TM: inserted] genome editing
    Article Snippet: Paragraph title: Southern blot ... The Kell-LPETG-targeted alleles were detected using anti-Dioxigenin-AP Fab fragment (Roche Diagnostics; 11093274910).

    RNA In Situ Hybridization:

    Article Title: Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
    Article Snippet: Paragraph title: RNA in situ hybridization ... Cells were then washed three times in 2× SSC and once in 4× SSC and incubated for 1 h at room temperature with 1:200 anti-digoxigenin-rhodamine Fab fragments (Cat. No. 11207750910, Roche Applied Science) in 4× SSC.

    Article Title: piRNAs and Aubergine cooperate with Wispy poly(A) polymerase to stabilize mRNAs in the germ plasm
    Article Snippet: Paragraph title: Immunostaining and RNA in situ hybridization ... Embryos were incubated with preabsorbed anti-Digoxigenin-alcaline phosphatase antibody (Roche, 11093274910) at 1:2000 for 1.5 h at room temperature on a wheel.

    In Vitro:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: PCR amplification of these plasmids using M13 forward and reverse primers produced products that included T7 and SP6 RNA polymerase binding sites which were used as templates for in vitro transcription to yield sense and antisense cRNAs using digoxigenin-(DIG-) labeled dNTPs (Roche #1277073). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Article Title:
    Article Snippet: The linearized template generated from EcoRI or NotI digest was used to generate digoxigenin-labeled sense and antisense probes by in vitro transcription using T3 and T7polymerase (Roche Applied Science catalog nos. .. The hybridized probe was detected after overnight incubation at 4 °C using an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche Applied Science catalog no. 11093274910).

    Northern Blot:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Paragraph title: RNA isolation, cDNA synthesis, northern blot analysis and in situ hybridization. ... Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Labeling:

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems
    Article Snippet: The digoxigenin and dinitrophenol labeled probes were simultaneously detected by incubation for 2 or more hours at RT (or at 4°C overnight) in a mixture of anti-digoxigenin POD fragments (Roche 11207733910) diluted 1:500 and anti-dinitrophenol AP conjugates 1:1000 (Vector laboratories MB-3100). .. Alternatively, embryos were incubated in a mixture of anti-digoxigenin AP FAB fragments (Roche 11093274910) diluted 1:4500 and anti-dinitrophenol POD (Perkin Elmer: Waltham, MA, USA TSA Plus DNP System NEL747A001KT) conjugates diluted 1:100.

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: PCR amplification of these plasmids using M13 forward and reverse primers produced products that included T7 and SP6 RNA polymerase binding sites which were used as templates for in vitro transcription to yield sense and antisense cRNAs using digoxigenin-(DIG-) labeled dNTPs (Roche #1277073). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Article Title: Small-molecule Wnt agonists correct cleft palates in Pax9 mutant mice in utero
    Article Snippet: In situ hybridizations were performed using digoxigenin-labeled RNA probes to Dkk1 , Dkk2 , Pax9 , Osr2 and Col1a1 as described previously ( ; ). .. 1 µg/ml antisense RNA probe was loaded on each section and anti-digoxigenin-AP antibody (11093274910, Roche; 1:1000) was used to detect the labeled probe. .. BM-purple (11442074001, Roche) was used for color development.

    Article Title: Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon
    Article Snippet: The purified Stx1B subunits were labeled with digoxigenin (DIG-Stx1B) as described previously ( ). .. Horseradish peroxidase (HRP)-labeled sheep anti-digoxigenin (DIG) Fab fragments (HRP-anti-DIG) were purchased from Roche Diagnostics (Tokyo, Japan).

    Generated:

    Article Title:
    Article Snippet: RNA probes were generated from the mouse IMAGE cDNA clone EMM1002-3872135 (Open Biosystems, Huntsville, AL), added to the hybridization buffer, hybridized to tissue sections overnight at 55 °C in a humidity chamber, and washed with 5×, 2×, and 0.2× SSC (1× SSC = 0.15 m NaCl and 0.015 m sodium citrate) buffer. .. The hybridized probe was detected after overnight incubation at 4 °C using an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche Applied Science catalog no. 11093274910).

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Chemiluminescent signal generated by CDP-Star (Roche, #11685672001) substrate was detected by exposure of membranes to Kodak Hyperfilm (Amersham Biosciences, #28-9068-36). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042).

    Sequencing:

    Article Title: Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
    Article Snippet: After three washes in phosphate-buffered saline cells were permeabilized by treatment with 0.1% triton in phosphate-buffered saline for 5 min and hybridized overnight at 37°C in 20 µl of a mixture containing 10% dextran sulphate, 10 µg yeast tRNA, 1× SSC, 20% formamide and 20 ng of digoxigenin-11-dUTP (Cat. No. 11573152910, Roche Applied Science) labelled PCR product probe against GFP coding sequence. .. Cells were then washed three times in 2× SSC and once in 4× SSC and incubated for 1 h at room temperature with 1:200 anti-digoxigenin-rhodamine Fab fragments (Cat. No. 11207750910, Roche Applied Science) in 4× SSC.

    Recombinant:

    Article Title: Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon
    Article Snippet: Recombinant binding subunits of Stx1 (Stx1B) were expressed in E. coli JM105(pVT1-B5) cells, and proteins were purified to homogeneity by anion-exchange chromatography and chromatofocusing as described previously ( ). .. Horseradish peroxidase (HRP)-labeled sheep anti-digoxigenin (DIG) Fab fragments (HRP-anti-DIG) were purchased from Roche Diagnostics (Tokyo, Japan).

    Isolation:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Paragraph title: RNA isolation, cDNA synthesis, northern blot analysis and in situ hybridization. ... Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Avidin-Biotin Assay:

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. Sections were deparaffinized by heating to 60 °C for 15 min, washed 3 × 5 min in xylene, rehydrated in a series of ethanol solutions (100%, 95%, 90% and 70% ethanol), washed in water and in 0.1% Tween-20 for 1 min each, steamed in antigen unmasking solution (Vector Laboratories H-3300) for 15 min, washed in phosphate-buffered saline containing 0.2% Tween-20 (PBST) for 5 min, blocked in 4% BSA in PBST for 30 min, incubated with rabbit anti-Sox9 antibodies (Millipore AB5535 at 1.7 μg ml−1 ) overnight at 4° in 1% BSA in PBST, washed three times for 5 min in PBST, incubated with Alexa 488-conjugated goat anti-rabbit antibody (Invitrogen A-11034 at 2.5 μg ml−1 ) for 2 hours, washed three times for five minutes in PBST including 0.2 μg ml−1 4,6-diamidino-2-phenylindole (DAPI) in the penultimate wash, and mounted in ProLong Gold antifade reagent (Thermo Fisher Scientific).

    Microscopy:

    Article Title: Small-molecule Wnt agonists correct cleft palates in Pax9 mutant mice in utero
    Article Snippet: 1 µg/ml antisense RNA probe was loaded on each section and anti-digoxigenin-AP antibody (11093274910, Roche; 1:1000) was used to detect the labeled probe. .. 1 µg/ml antisense RNA probe was loaded on each section and anti-digoxigenin-AP antibody (11093274910, Roche; 1:1000) was used to detect the labeled probe.

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042).

    Article Title: Fog1 is required for cardiac looping in zebrafish
    Article Snippet: An alkaline phosphatase conjugated sheep anti-digoxigenin Fab fragment (Roche) was added (1:5000 dilution) and incubated for 16 h at 4°C. .. An alkaline phosphatase conjugated sheep anti-digoxigenin Fab fragment (Roche) was added (1:5000 dilution) and incubated for 16 h at 4°C.

    Purification:

    Article Title: Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon
    Article Snippet: The purified Stx1B subunits were labeled with digoxigenin (DIG-Stx1B) as described previously ( ). .. Horseradish peroxidase (HRP)-labeled sheep anti-digoxigenin (DIG) Fab fragments (HRP-anti-DIG) were purchased from Roche Diagnostics (Tokyo, Japan).

    Polymerase Chain Reaction:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: PCR amplification of these plasmids using M13 forward and reverse primers produced products that included T7 and SP6 RNA polymerase binding sites which were used as templates for in vitro transcription to yield sense and antisense cRNAs using digoxigenin-(DIG-) labeled dNTPs (Roche #1277073). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Article Title: Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
    Article Snippet: After three washes in phosphate-buffered saline cells were permeabilized by treatment with 0.1% triton in phosphate-buffered saline for 5 min and hybridized overnight at 37°C in 20 µl of a mixture containing 10% dextran sulphate, 10 µg yeast tRNA, 1× SSC, 20% formamide and 20 ng of digoxigenin-11-dUTP (Cat. No. 11573152910, Roche Applied Science) labelled PCR product probe against GFP coding sequence. .. Cells were then washed three times in 2× SSC and once in 4× SSC and incubated for 1 h at room temperature with 1:200 anti-digoxigenin-rhodamine Fab fragments (Cat. No. 11207750910, Roche Applied Science) in 4× SSC.

    Immunostaining:

    Article Title: piRNAs and Aubergine cooperate with Wispy poly(A) polymerase to stabilize mRNAs in the germ plasm
    Article Snippet: Paragraph title: Immunostaining and RNA in situ hybridization ... Embryos were incubated with preabsorbed anti-Digoxigenin-alcaline phosphatase antibody (Roche, 11093274910) at 1:2000 for 1.5 h at room temperature on a wheel.

    Mouse Assay:

    Article Title: Silencing microRNA-134 produces neuroprotective and prolonged seizure-suppressive effects
    Article Snippet: Mice (n = 3) were perfused with 4% paraformaldehyde and 12 μm tissue sections mounted on SuperFrost-Plus slides (VWR). .. Then sections were rinsed in B1 buffer (150 mM NaCl, 100 mM Maleic acid and 0.4% IGEPAL pH:7.5) for 1 h at room temperature and B2 buffer (2% blocking reagent and 10% goat serum in B1) for 30 min. Polyclonal antibodies against digoxigenin (1:1000, Cat. 11093274910, Roche) were incubated in B2 overnight at 4 °C.

    In Situ Hybridization:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome
    Article Snippet: Paragraph title: 2.3. In situ hybridization ... Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Article Title: Differential effects of acute and chronic ethanol exposure on orexin expression in the perifornical lateral hypothalamus
    Article Snippet: Paragraph title: Digoxigenin-labeled In Situ Hybridization Histochemistry ... AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000, Roche, Nutley, NJ) and NBT / BCIP (Roche, Nutley, NJ) were used to visualize the signal.

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Paragraph title: RNA isolation, cDNA synthesis, northern blot analysis and in situ hybridization. ... Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Article Title:
    Article Snippet: Paragraph title: In Situ Hybridization ... The hybridized probe was detected after overnight incubation at 4 °C using an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche Applied Science catalog no. 11093274910).

    Article Title: PRENATAL ETHANOL EXPOSURE STIMULATES NEUROGENESIS IN HYPOTHALAMIC AND LIMBIC PEPTIDE SYSTEMS: POSSIBLE MECHANISM FOR OFFSPRING ETHANOL OVERCONSUMPTION
    Article Snippet: Paragraph title: Digoxigenin-labeled in situ hybridization histochemistry ... AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000; Roche Diagnostics, Indianapolis, IN, USA) and NBT/BCIP (Roche Diagnostics) were used to visualize the signal.

    Article Title: Fog1 is required for cardiac looping in zebrafish
    Article Snippet: Paragraph title: In situ hybridization ... An alkaline phosphatase conjugated sheep anti-digoxigenin Fab fragment (Roche) was added (1:5000 dilution) and incubated for 16 h at 4°C.

    Article Title: Glutamatergic input to the lateral hypothalamus stimulates ethanol intake: Role of orexin and melanin-concentrating hormone
    Article Snippet: Paragraph title: Digoxigenin-Labeled In Situ Hybridization Histochemistry ... AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000, Roche, Nutley, NJ) and NBT/BCIP (Roche, Nutley, NJ) were used to visualize the signal.

    Article Title: piRNAs and Aubergine cooperate with Wispy poly(A) polymerase to stabilize mRNAs in the germ plasm
    Article Snippet: For whole-mount in situ hybridization experiments, fixed embryos were rehydrated gradually with methanol-PBT-Tween (PBS supplemented with 0.1% Tween-20) and washed three times in PBT-Tween. .. Embryos were incubated with preabsorbed anti-Digoxigenin-alcaline phosphatase antibody (Roche, 11093274910) at 1:2000 for 1.5 h at room temperature on a wheel.

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: Briefly, for in situ hybridization, formalin-fixed and paraffin-embedded samples were deparaffinized, rehydrated, treated with HCl, digested in proteinase K solution, refixed with formaldehyde, treated in acetic anhydride solution, and hybridized with a digoxigenin labelled RNA probe specific for Ascl2 . .. After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution.

    Plasmid Preparation:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome
    Article Snippet: Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C. .. Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems
    Article Snippet: The digoxigenin probe was detected by incubation for 2 or more hours at RT (or at 4°C overnight) with anti-digoxigenin FAB fragments (Roche Scandinavia: Bromma, Sweden 11093274910) conjugated to AP diluted 1:4500. .. The digoxigenin probe was detected by incubation for 2 or more hours at RT (or at 4°C overnight) with anti-digoxigenin FAB fragments (Roche Scandinavia: Bromma, Sweden 11093274910) conjugated to AP diluted 1:4500.

    Article Title:
    Article Snippet: The resulting SLIP-GC vector was linearized by digestion with either EcoRI or NotI restriction enzymes. .. The hybridized probe was detected after overnight incubation at 4 °C using an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche Applied Science catalog no. 11093274910).

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution.

    Software:

    Article Title: PRENATAL ETHANOL EXPOSURE STIMULATES NEUROGENESIS IN HYPOTHALAMIC AND LIMBIC PEPTIDE SYSTEMS: POSSIBLE MECHANISM FOR OFFSPRING ETHANOL OVERCONSUMPTION
    Article Snippet: AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000; Roche Diagnostics, Indianapolis, IN, USA) and NBT/BCIP (Roche Diagnostics) were used to visualize the signal. .. Sections were viewed on a Leitz microscope (10× objective).

    Negative Control:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042).

    Binding Assay:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: PCR amplification of these plasmids using M13 forward and reverse primers produced products that included T7 and SP6 RNA polymerase binding sites which were used as templates for in vitro transcription to yield sense and antisense cRNAs using digoxigenin-(DIG-) labeled dNTPs (Roche #1277073). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Article Title: Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon
    Article Snippet: Recombinant binding subunits of Stx1 (Stx1B) were expressed in E. coli JM105(pVT1-B5) cells, and proteins were purified to homogeneity by anion-exchange chromatography and chromatofocusing as described previously ( ). .. Horseradish peroxidase (HRP)-labeled sheep anti-digoxigenin (DIG) Fab fragments (HRP-anti-DIG) were purchased from Roche Diagnostics (Tokyo, Japan).

    Agarose Gel Electrophoresis:

    Article Title: Inhibition of non-homologous end joining increases the efficiency of CRISPR/Cas9-mediated precise [TM: inserted] genome editing
    Article Snippet: Genomic DNA was separated on a 0.5% agarose gel after digesting with EcoRI restriction enzyme, transferred to a nylon membrane (Millipore; INYC00010) and hybridized with PCRDIG (Roche Diagnostics; 11636090910)-labeled probes. .. The Kell-LPETG-targeted alleles were detected using anti-Dioxigenin-AP Fab fragment (Roche Diagnostics; 11093274910).

    Chromatography:

    Article Title: Restricted Expression of Shiga Toxin Binding Sites on Mucosal Epithelium of Mouse Distal Colon
    Article Snippet: Recombinant binding subunits of Stx1 (Stx1B) were expressed in E. coli JM105(pVT1-B5) cells, and proteins were purified to homogeneity by anion-exchange chromatography and chromatofocusing as described previously ( ). .. Horseradish peroxidase (HRP)-labeled sheep anti-digoxigenin (DIG) Fab fragments (HRP-anti-DIG) were purchased from Roche Diagnostics (Tokyo, Japan).

    Produced:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: PCR amplification of these plasmids using M13 forward and reverse primers produced products that included T7 and SP6 RNA polymerase binding sites which were used as templates for in vitro transcription to yield sense and antisense cRNAs using digoxigenin-(DIG-) labeled dNTPs (Roche #1277073). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910).

    Concentration Assay:

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042). .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042).

    Article Title: Fog1 is required for cardiac looping in zebrafish
    Article Snippet: An alkaline phosphatase conjugated sheep anti-digoxigenin Fab fragment (Roche) was added (1:5000 dilution) and incubated for 16 h at 4°C. .. Following this incubation, embryos were washed 10 times with PBST for 15 min each at room temperature and then 3 times for 5 min each with AP buffer (100 mM Tris pH 9.5, 50 mM MgCl2, 100 mM NaCl, 0.1% Tween-20).

    In Situ:

    Article Title: Small-molecule Wnt agonists correct cleft palates in Pax9 mutant mice in utero
    Article Snippet: Paragraph title: Microscopic evaluations and in situ hybridizations ... 1 µg/ml antisense RNA probe was loaded on each section and anti-digoxigenin-AP antibody (11093274910, Roche; 1:1000) was used to detect the labeled probe.

    Article Title: Active nuclear transcriptome analysis reveals inflammasome-dependent mechanism for early neutrophil response to Mycobacterium marinum
    Article Snippet: Paragraph title: Whole-mount in situ hybridisations ... Probes were detected with an anti-DIG-alkaline phosphatase antibody (Roche, cat # 11093274910) and the signal was developed using BM Purple AP (Roche, cat # 11442074001).

    Article Title: PRENATAL ETHANOL EXPOSURE STIMULATES NEUROGENESIS IN HYPOTHALAMIC AND LIMBIC PEPTIDE SYSTEMS: POSSIBLE MECHANISM FOR OFFSPRING ETHANOL OVERCONSUMPTION
    Article Snippet: For better visualization, in situ hydridization histochemistry with a digoxigenin-labeled probe (digoxigenin-labeled in situ hybridization; DIG-ISH) was performed to reveal the anatomical distribution at birth (P0) of changes in peptide expression in the control, 1 g/kg ethanol and 3 g/kg dose ethanol groups ( n = 6–8/group) as described ( , ). .. AP-conjugated sheep anti-digoxigenin Fab fragments (1:1000; Roche Diagnostics, Indianapolis, IN, USA) and NBT/BCIP (Roche Diagnostics) were used to visualize the signal.

    Article Title: Silencing microRNA-134 produces neuroprotective and prolonged seizure-suppressive effects
    Article Snippet: Paragraph title: In situ hybridization ... Then sections were rinsed in B1 buffer (150 mM NaCl, 100 mM Maleic acid and 0.4% IGEPAL pH:7.5) for 1 h at room temperature and B2 buffer (2% blocking reagent and 10% goat serum in B1) for 30 min. Polyclonal antibodies against digoxigenin (1:1000, Cat. 11093274910, Roche) were incubated in B2 overnight at 4 °C.

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. For immunohistochemical detection of Ki67, sections were deparaffinized, rehydrated and subjected to antigen unmasking in the same fashion as for Sox9 staining, and were then treated with 2.25% hydrogen peroxide for 15 min, washed in water and PBST for 5 min each, blocked with Avidin D reagent (Vector Laboratories SP-2001), washed in PBST, blocked with 4% BSA in PBST for 30 min, incubated with rabbit anti-Ki67 antibody (Abcam Ab16667 at 1:100), washed three times for 5 min in PBST, incubated with biotinylated anti-rabbit antibody (Vector Laboratories BA-1000 at 7.5 μg ml−1 ) for 30 min, washed in PBST 3–5 min, stained with the Vector Laboratories ABC and DAB reagents (PK-6200 and SK-4100), counterstained with Gill #2 haematoxylin (Thermo Fisher Scientific), dehydrated in an ethanol series (70–100%), cleared in xylene, and mounted in Cytoseal XYL (Thermo Fisher Scientific 8312-4).

    Staining:

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems
    Article Snippet: Alternatively, embryos were incubated in a mixture of anti-digoxigenin AP FAB fragments (Roche 11093274910) diluted 1:4500 and anti-dinitrophenol POD (Perkin Elmer: Waltham, MA, USA TSA Plus DNP System NEL747A001KT) conjugates diluted 1:100. .. For visualization of the two different transcript probes, the POD-TSA reaction was performed first followed by AP-Fast Red or AP-Fast Blue staining.

    Article Title: Small-molecule Wnt agonists correct cleft palates in Pax9 mutant mice in utero
    Article Snippet: 1 µg/ml antisense RNA probe was loaded on each section and anti-digoxigenin-AP antibody (11093274910, Roche; 1:1000) was used to detect the labeled probe. .. 1 µg/ml antisense RNA probe was loaded on each section and anti-digoxigenin-AP antibody (11093274910, Roche; 1:1000) was used to detect the labeled probe.

    Article Title: TGF? superfamily signaling regulators are differentially expressed in the developing and adult mouse testis
    Article Snippet: Hybridization was performed with 100–400 ng probe per slide at 50–60°C with stringency washes to 0.1× SSC at the hybridization temperature. .. Bound DIG-labeled riboprobe was detected using an anti-DIG antibody (Roche, #11093274910) and visualized by purple staining using 5-Bromo-4chloro-3-indoyl phosphate/nitroblue tetrazolium (BCIP/NBT) substrate (Thermo Scientific, #34042). .. Sections were counterstained with Harris haematoxylin to visualize chromatin and mounted in GVA aqueous mounting solution (Invitrogen #00-800).

    Article Title: Fog1 is required for cardiac looping in zebrafish
    Article Snippet: An alkaline phosphatase conjugated sheep anti-digoxigenin Fab fragment (Roche) was added (1:5000 dilution) and incubated for 16 h at 4°C. .. Following this incubation, embryos were washed 10 times with PBST for 15 min each at room temperature and then 3 times for 5 min each with AP buffer (100 mM Tris pH 9.5, 50 mM MgCl2, 100 mM NaCl, 0.1% Tween-20).

    Article Title: Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche
    Article Snippet: After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution. .. After washes with 2 × saline-sodium citrate (SSC) and 50% formamide/2 × SSC, sections were incubated overnight at 4 °C with alkaline phosphatase-conjugated anti-digoxigenin antibodies (Roche #11093274910 at a 1:2,000 dilution, that is, 0.375 units per ml), and developed using nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) solution.

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    Roche sheep anti digoxigenin fab fragments antibody ap conjugated
    Representative images from a female mouse at weaning age to illustrate <t>digoxigenin-based</t> Snord116 in situ hybridization (ISH) signal in multiple brain regions. Panels A (caudal diencephalon) and B (rostral mesencephalon) illustrate coronal sections hundreds
    Sheep Anti Digoxigenin Fab Fragments Antibody Ap Conjugated, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 150 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sheep anti digoxigenin fab fragments antibody ap conjugated/product/Roche
    Average 99 stars, based on 150 article reviews
    Price from $9.99 to $1999.99
    sheep anti digoxigenin fab fragments antibody ap conjugated - by Bioz Stars, 2019-10
    99/100 stars
      Buy from Supplier

    99
    Roche sheep anti digoxigenin fab fragments antibody pod conjugated
    Inactivation of <t>antibody-POD</t> conjugate . Zebrafish embryos at 28 hpf were hybridized with dinitrophenyl-labeled shha and <t>digoxigenin-labeled</t> nkx6.1 RNA probes. (A, B) The expression patterns of shha (A) and nkx6.1 (B) as seen in single-color FISH experiments. In two-color experiments shha transcript was detected first using DyLight633-tyramide and nkx6.1 transcript was detected subsequently by FAM-tyramide. (C-H) Prior to the second round of detection, embryos were incubated for 10 minutes in PBS T (PBS plus 0.1% Tween-20) (C, D), PBS T containing 6% H 2 O 2 (E, F), or 100 mM glycine-HCl pH 2.0 (G, H). Single confocal sections of zebrafish brains are shown in the DyLight633-detection channel (Ch01) and in the FAM-detection channel (Ch02) from a lateral view and with anterior to the left. Images were recorded on a LSM510 microscope (Carl Zeiss) and false colored in ImageJ. Scale bar = 50 μm.
    Sheep Anti Digoxigenin Fab Fragments Antibody Pod Conjugated, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sheep anti digoxigenin fab fragments antibody pod conjugated/product/Roche
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    sheep anti digoxigenin fab fragments antibody pod conjugated - by Bioz Stars, 2019-10
    99/100 stars
      Buy from Supplier

    Image Search Results


    Representative images from a female mouse at weaning age to illustrate digoxigenin-based Snord116 in situ hybridization (ISH) signal in multiple brain regions. Panels A (caudal diencephalon) and B (rostral mesencephalon) illustrate coronal sections hundreds

    Journal:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome

    doi: 10.1016/j.ijdevneu.2012.05.005

    Figure Lengend Snippet: Representative images from a female mouse at weaning age to illustrate digoxigenin-based Snord116 in situ hybridization (ISH) signal in multiple brain regions. Panels A (caudal diencephalon) and B (rostral mesencephalon) illustrate coronal sections hundreds

    Article Snippet: Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Techniques: In Situ Hybridization

    Representative images from a female mouse at weaning age show digoxigenin-based Snord116 in situ hybridization (ISH) signals in the hypothalamus. The ISH signal was detected in the paraventricular nucleus (PVN; panel A), ventromedial nucleus of the hypothalamus

    Journal:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome

    doi: 10.1016/j.ijdevneu.2012.05.005

    Figure Lengend Snippet: Representative images from a female mouse at weaning age show digoxigenin-based Snord116 in situ hybridization (ISH) signals in the hypothalamus. The ISH signal was detected in the paraventricular nucleus (PVN; panel A), ventromedial nucleus of the hypothalamus

    Article Snippet: Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Techniques: In Situ Hybridization

    Representative digoxigenin-based in situ hybridization (ISH) images of Snord116 from the rostral to caudal hypothalamus in a female mouse at weaning age. The images from panels A to L are hypothalamic sections taken from rostral to caudal. The Snord116

    Journal:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome

    doi: 10.1016/j.ijdevneu.2012.05.005

    Figure Lengend Snippet: Representative digoxigenin-based in situ hybridization (ISH) images of Snord116 from the rostral to caudal hypothalamus in a female mouse at weaning age. The images from panels A to L are hypothalamic sections taken from rostral to caudal. The Snord116

    Article Snippet: Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Techniques: In Situ Hybridization

    Digital images show digoxigenin-based in situ hybridization (ISH) illustrating the expression of Snord116 in the brain including hypothalamus and surrounding area in neonatal (P0) mice. Panels A and B show the representative images of Snord116 in mice

    Journal:

    Article Title: Hypothalamic Expression of SnoRNA Snord116 is Consistent with a Link to the hyperphagia and obesity symptoms of Prader-Willi Syndrome

    doi: 10.1016/j.ijdevneu.2012.05.005

    Figure Lengend Snippet: Digital images show digoxigenin-based in situ hybridization (ISH) illustrating the expression of Snord116 in the brain including hypothalamus and surrounding area in neonatal (P0) mice. Panels A and B show the representative images of Snord116 in mice

    Article Snippet: Sections then were incubated with 1:2000 dilution of alkaline phosphatase conjugated anti-DIG antibody (Roche, 11093274910) in 1% sheep serum/TBST overnight at 4°C.

    Techniques: In Situ Hybridization, Expressing, Mouse Assay

    Control of bleed-through between Fast Blue and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Blue (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Blue (Ch01: detection of wavelengths greater than 650 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. No bleed-through was detected between the TSA-FAM and Fast Blue detection channels (B,C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Journal: BMC Developmental Biology

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems

    doi: 10.1186/1471-213X-11-43

    Figure Lengend Snippet: Control of bleed-through between Fast Blue and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Blue (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Blue (Ch01: detection of wavelengths greater than 650 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. No bleed-through was detected between the TSA-FAM and Fast Blue detection channels (B,C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Article Snippet: The digoxigenin probe was detected by incubation for 2 or more hours at RT (or at 4°C overnight) with anti-digoxigenin FAB fragments (Roche Scandinavia: Bromma, Sweden 11093274910) conjugated to AP diluted 1:4500.

    Techniques: Labeling, Fluorescence, Microscopy

    Fast Red and Fast Blue permit chromogenic and fluorescent transcript visualization . Lateral views of embryonic brains at 24 hpf hybridized to nkx6.1 digoxigenin and/or pax6a dinitrophenol antisense RNA probes as indicated on each panel. Transcript distributions were visualized by chromogenic (A,C,E) or fluorescent (B,D) detection of Fast Blue (C,D,E) and Fast Red (A,B,E) precipitates as indicated. Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Fluorescent signals were false-colored in ImageJ. Scale bar is 100 μm.

    Journal: BMC Developmental Biology

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems

    doi: 10.1186/1471-213X-11-43

    Figure Lengend Snippet: Fast Red and Fast Blue permit chromogenic and fluorescent transcript visualization . Lateral views of embryonic brains at 24 hpf hybridized to nkx6.1 digoxigenin and/or pax6a dinitrophenol antisense RNA probes as indicated on each panel. Transcript distributions were visualized by chromogenic (A,C,E) or fluorescent (B,D) detection of Fast Blue (C,D,E) and Fast Red (A,B,E) precipitates as indicated. Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Fluorescent signals were false-colored in ImageJ. Scale bar is 100 μm.

    Article Snippet: The digoxigenin probe was detected by incubation for 2 or more hours at RT (or at 4°C overnight) with anti-digoxigenin FAB fragments (Roche Scandinavia: Bromma, Sweden 11093274910) conjugated to AP diluted 1:4500.

    Techniques: Microscopy

    Control of bleed-through between Fast Red and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Red (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Red (Ch01: detection of wavelengths greater than 560 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. A significant bleed-through of TSA-FAM nkx6.1 signal was detected in the Fast Red detection channel (C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Journal: BMC Developmental Biology

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems

    doi: 10.1186/1471-213X-11-43

    Figure Lengend Snippet: Control of bleed-through between Fast Red and TSA-FAM detection channels . 24-hpf embryos were hybridized to a dinitrophenol-labeled pax6a probe (A,B) or to a digoxigenin-labeled antisense RNA probe specific for nkx6.1 (C,D). Transcripts were detected using Fast Red (A,B) and TSA-FAM (C,D). Fluorescence signals were recorded in the Fast Red (Ch01: detection of wavelengths greater than 560 nm) and TSA-FAM (Ch02: detection of wavelengths from 505 nm to 545 nm) detection channels. A significant bleed-through of TSA-FAM nkx6.1 signal was detected in the Fast Red detection channel (C). Lateral views with anterior to the left are shown. Images were recorded on a LSM510 confocal microscope and false-colored in ImageJ. Scale bar is 50 μm.

    Article Snippet: The digoxigenin probe was detected by incubation for 2 or more hours at RT (or at 4°C overnight) with anti-digoxigenin FAB fragments (Roche Scandinavia: Bromma, Sweden 11093274910) conjugated to AP diluted 1:4500.

    Techniques: Labeling, Fluorescence, Microscopy

    Effects of dextran sulfate in standard WISH . 24-hpf embryos were hybridized to a sim1a digoxigenin-labeled RNA probe, which was visualized by AP-BCIP/NBT chromogenic detection under identical conditions and staining times. Lateral views of embryonic brains (A,B) and trunks (C,D) are shown with anterior to the left. In (B,D) but not in (A,C) 5% dextran sulfate was included in the hybridization buffer. Addition of dextran sulfate resulted in increased signal sensitivity. Arrowheads in (A,B) indicate sim1a -positive neuronal clusters identified in dextran sulfate treated brains (B) that were hardly detected in untreated embryos (A). Arrowheads in (C,D) mark the pronephric primordium strongly visualized in dextrane sulfate treated embryos (D) but not in untreated specimens (C). Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Scale bar is 100 μm.

    Journal: BMC Developmental Biology

    Article Title: Two-color fluorescent in situ hybridization in the embryonic zebrafish brain using differential detection systems

    doi: 10.1186/1471-213X-11-43

    Figure Lengend Snippet: Effects of dextran sulfate in standard WISH . 24-hpf embryos were hybridized to a sim1a digoxigenin-labeled RNA probe, which was visualized by AP-BCIP/NBT chromogenic detection under identical conditions and staining times. Lateral views of embryonic brains (A,B) and trunks (C,D) are shown with anterior to the left. In (B,D) but not in (A,C) 5% dextran sulfate was included in the hybridization buffer. Addition of dextran sulfate resulted in increased signal sensitivity. Arrowheads in (A,B) indicate sim1a -positive neuronal clusters identified in dextran sulfate treated brains (B) that were hardly detected in untreated embryos (A). Arrowheads in (C,D) mark the pronephric primordium strongly visualized in dextrane sulfate treated embryos (D) but not in untreated specimens (C). Embryos were viewed on an Axioplan II microscope and images were recorded with an Axiocam digital camera. Scale bar is 100 μm.

    Article Snippet: The digoxigenin probe was detected by incubation for 2 or more hours at RT (or at 4°C overnight) with anti-digoxigenin FAB fragments (Roche Scandinavia: Bromma, Sweden 11093274910) conjugated to AP diluted 1:4500.

    Techniques: Labeling, Staining, Hybridization, Microscopy

    Inactivation of antibody-POD conjugate . Zebrafish embryos at 28 hpf were hybridized with dinitrophenyl-labeled shha and digoxigenin-labeled nkx6.1 RNA probes. (A, B) The expression patterns of shha (A) and nkx6.1 (B) as seen in single-color FISH experiments. In two-color experiments shha transcript was detected first using DyLight633-tyramide and nkx6.1 transcript was detected subsequently by FAM-tyramide. (C-H) Prior to the second round of detection, embryos were incubated for 10 minutes in PBS T (PBS plus 0.1% Tween-20) (C, D), PBS T containing 6% H 2 O 2 (E, F), or 100 mM glycine-HCl pH 2.0 (G, H). Single confocal sections of zebrafish brains are shown in the DyLight633-detection channel (Ch01) and in the FAM-detection channel (Ch02) from a lateral view and with anterior to the left. Images were recorded on a LSM510 microscope (Carl Zeiss) and false colored in ImageJ. Scale bar = 50 μm.

    Journal: Neural Development

    Article Title: Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

    doi: 10.1186/1749-8104-6-10

    Figure Lengend Snippet: Inactivation of antibody-POD conjugate . Zebrafish embryos at 28 hpf were hybridized with dinitrophenyl-labeled shha and digoxigenin-labeled nkx6.1 RNA probes. (A, B) The expression patterns of shha (A) and nkx6.1 (B) as seen in single-color FISH experiments. In two-color experiments shha transcript was detected first using DyLight633-tyramide and nkx6.1 transcript was detected subsequently by FAM-tyramide. (C-H) Prior to the second round of detection, embryos were incubated for 10 minutes in PBS T (PBS plus 0.1% Tween-20) (C, D), PBS T containing 6% H 2 O 2 (E, F), or 100 mM glycine-HCl pH 2.0 (G, H). Single confocal sections of zebrafish brains are shown in the DyLight633-detection channel (Ch01) and in the FAM-detection channel (Ch02) from a lateral view and with anterior to the left. Images were recorded on a LSM510 microscope (Carl Zeiss) and false colored in ImageJ. Scale bar = 50 μm.

    Article Snippet: Sheep-anti-digoxigenin-POD Fab fragments (Roche 11207733910) were diluted 1:500, anti-dinitrophenyl-POD (PerkinElmer TSA Plus DNP System NEL747A001KT) was diluted 1:100 and the rabbit-anti-fluorescein/Oregon Green 488-POD (Molecular Probes: Eugene, OR, USA A21253) was diluted 1:500.

    Techniques: Labeling, Expressing, Fluorescence In Situ Hybridization, Incubation, Microscopy

    Positive effects of substituted phenol compounds on the TSA-POD reaction . Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for nkx6.1 are shown with anterior to the left. (A) Control (CTL); (B, C) vanillin added; (D, E) 4-iodophenol added. In all embryos shown, dextran sulfate was included in the hybridization and TSA-POD reaction. Above each panel signal-to-noise ratios (s/n) and compound concentrations are indicated. Black and white pictures were recorded with identical exposure times. Images were false-colored with help of the ImageJ software and no further adjustments or other image processing were performed. Scale bar = 100 μm.

    Journal: Neural Development

    Article Title: Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

    doi: 10.1186/1749-8104-6-10

    Figure Lengend Snippet: Positive effects of substituted phenol compounds on the TSA-POD reaction . Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for nkx6.1 are shown with anterior to the left. (A) Control (CTL); (B, C) vanillin added; (D, E) 4-iodophenol added. In all embryos shown, dextran sulfate was included in the hybridization and TSA-POD reaction. Above each panel signal-to-noise ratios (s/n) and compound concentrations are indicated. Black and white pictures were recorded with identical exposure times. Images were false-colored with help of the ImageJ software and no further adjustments or other image processing were performed. Scale bar = 100 μm.

    Article Snippet: Sheep-anti-digoxigenin-POD Fab fragments (Roche 11207733910) were diluted 1:500, anti-dinitrophenyl-POD (PerkinElmer TSA Plus DNP System NEL747A001KT) was diluted 1:100 and the rabbit-anti-fluorescein/Oregon Green 488-POD (Molecular Probes: Eugene, OR, USA A21253) was diluted 1:500.

    Techniques: Labeling, CTL Assay, Hybridization, Software

    Bench-made fluorescent tyramides applied in zebrafish whole-mount FISH . Embryos were viewed on an Axioplan II microscope (Carl Zeiss). For excitation, a mercury burner (HBO 103 OSRAM) was used. Lateral views of zebrafish brains are shown with anterior to the left. Scale bar = 100 μm. ( A-C ) Zebrafish embryos at 28 hpf hybridized to a digoxigenin-labeled nkx6.1 antisense RNA probe. Images were captured with an Orca digital camera (Hamamatsu). As POD substrates, three different bench-made fluorogenic tyramides at a 1:250 dilution were used: (A) FAM-tyramide (exposure time, 100 ms; Chroma-filter 41001); (B) TAMRA-tyramide (exposure time, 4 ms; Chroma-filter 41002b); and (C) DyLight633-tyramide (exposure time, 120 ms; Chroma-filter 41008). ( D-F ) Zebrafish embryos at 24 hpf hybridized to a digoxigenin-labeled shha antisense RNA probe. Images were recorded with an Axiocam digital color camera (Carl Zeiss) using identical exposure times. Transcript distribution was visualized with (D) fluorescein-tyramide from Perkin Elmer (SAT701B001EA) at a 1:100 dilution, (E) bench-made FAM-tyramide at a 1:250 dilution, or (F) a 1:100 dilution.

    Journal: Neural Development

    Article Title: Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

    doi: 10.1186/1749-8104-6-10

    Figure Lengend Snippet: Bench-made fluorescent tyramides applied in zebrafish whole-mount FISH . Embryos were viewed on an Axioplan II microscope (Carl Zeiss). For excitation, a mercury burner (HBO 103 OSRAM) was used. Lateral views of zebrafish brains are shown with anterior to the left. Scale bar = 100 μm. ( A-C ) Zebrafish embryos at 28 hpf hybridized to a digoxigenin-labeled nkx6.1 antisense RNA probe. Images were captured with an Orca digital camera (Hamamatsu). As POD substrates, three different bench-made fluorogenic tyramides at a 1:250 dilution were used: (A) FAM-tyramide (exposure time, 100 ms; Chroma-filter 41001); (B) TAMRA-tyramide (exposure time, 4 ms; Chroma-filter 41002b); and (C) DyLight633-tyramide (exposure time, 120 ms; Chroma-filter 41008). ( D-F ) Zebrafish embryos at 24 hpf hybridized to a digoxigenin-labeled shha antisense RNA probe. Images were recorded with an Axiocam digital color camera (Carl Zeiss) using identical exposure times. Transcript distribution was visualized with (D) fluorescein-tyramide from Perkin Elmer (SAT701B001EA) at a 1:100 dilution, (E) bench-made FAM-tyramide at a 1:250 dilution, or (F) a 1:100 dilution.

    Article Snippet: Sheep-anti-digoxigenin-POD Fab fragments (Roche 11207733910) were diluted 1:500, anti-dinitrophenyl-POD (PerkinElmer TSA Plus DNP System NEL747A001KT) was diluted 1:100 and the rabbit-anti-fluorescein/Oregon Green 488-POD (Molecular Probes: Eugene, OR, USA A21253) was diluted 1:500.

    Techniques: Fluorescence In Situ Hybridization, Microscopy, Labeling, Mass Spectrometry

    Optimization of hybridization and POD reaction by dextran sulfate . (A, C, E, G) Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for dbx1a are shown with anterior to the left. Above each panel signal-to-noise ratios (s/n) are indicated and whether dextran sulfate was added (+) or not (-) to the hybridization buffer (Hyb) and/or TSA reaction. (B, D, F, H) Embryos hybridized without probe were used as negative controls to assess background noise. Black-and-white pictures were recorded with identical exposure times using an Orca digital camera (Hamamatsu) on an Axioplan II microscope (Carl Zeiss). Images were false-colored with help of the ImageJ software and no further adjustments or other image processing was performed. Scale bar = 100 μm.

    Journal: Neural Development

    Article Title: Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

    doi: 10.1186/1749-8104-6-10

    Figure Lengend Snippet: Optimization of hybridization and POD reaction by dextran sulfate . (A, C, E, G) Lateral views of 1-dpf zebrafish embryos hybridized with a digoxigenin-labeled antisense RNA probe specific for dbx1a are shown with anterior to the left. Above each panel signal-to-noise ratios (s/n) are indicated and whether dextran sulfate was added (+) or not (-) to the hybridization buffer (Hyb) and/or TSA reaction. (B, D, F, H) Embryos hybridized without probe were used as negative controls to assess background noise. Black-and-white pictures were recorded with identical exposure times using an Orca digital camera (Hamamatsu) on an Axioplan II microscope (Carl Zeiss). Images were false-colored with help of the ImageJ software and no further adjustments or other image processing was performed. Scale bar = 100 μm.

    Article Snippet: Sheep-anti-digoxigenin-POD Fab fragments (Roche 11207733910) were diluted 1:500, anti-dinitrophenyl-POD (PerkinElmer TSA Plus DNP System NEL747A001KT) was diluted 1:100 and the rabbit-anti-fluorescein/Oregon Green 488-POD (Molecular Probes: Eugene, OR, USA A21253) was diluted 1:500.

    Techniques: Hybridization, Labeling, Microscopy, Software