Structured Review

Jackson Immuno alexa fluor 594
Arabidopsis HMGR localizes in the ER network, nuclear envelope, HMGR vesicles and ER-HMGR domains. ( a – c ) Whole-mount immunohistochemical analysis of cotyledon parenchymal cells from 6-day-old Arabidopsis WT seedlings ( a ) Immunodetection of HMGR with Ab-CD1-i and anti-rabbit IgG secondary antibody <t>(Alexa</t> Fluor 555, in red), visualized by confocal microscopy under dark (left) or bright fields (right). ( b ) Immunodetection of HMGR with Ab-CD1-i and anti-rabbit IgG secondary antibody (AlexaFluor 555, in red), and simultaneous detection of chlorophyll (in blue). ( c ) Negative control without the Ab-CD1-i antibody. The irregular corpuscles, 0.2 to 2 µm in length, correspond to HMGR vesicles. The elliptic bodies, 6 to 8 µm in diameter, correspond to chloroplasts. Images were obtained by Z-projection encompassing 3 ( a ), 10 ( b ) or 4 ( c ) µm in the Z-axis. Bars, 5 µm. ( d – f ) Immunochemical study of HMGR vesicles by transmission EM. Leaf samples from 10-day-old Arabidopsis WT or 1S:GFP seedlings were processed by HPF and embedded with Lowicryl HM20. ( d ) HMGR was detected in cotyledon from WT seedlings with Ab-CD1-i and anti-rabbit-IgG (18 nm particle). ( e ) HMGR was detected in true leaf from 1S:GFP seedlings with Ab-CD1-i and anti-rabbit-IgG (12 nm particle). ( f ) Double immunolocalization of HMGR and 1S:GFP in true leaf from 1S:GFP seedlings. HMGR was detected with Ab-CD1-i and anti-rabbit-IgG (12 nm particle) and 1S:GFP was detected with Ab-5450 and anti-goat-IgG (18 nm particle). Black and blue arrowheads indicate, respectively, the external and internal membranes from HMGR vesicles. Red arrowheads indicate ER strands. Bars, 250 nm. ( g ) Whole-mount immunohistochemical analysis of cotyledon parenchymal cells from 6-day-old Arabidopsis 1S:GFP transgenic seedlings. 1S:GFP was detected with Ab-5450 and secondary antibody Alexa fluor 488 (green). HMGR was detected with Ab-CD1-i and secondary antibody Alexa fluor 594 (red). Images were obtained by Z-projection encompassing 12 µm in the Z-axis. Bar, 5 µm. ( h – m ) Immunolocalization of HMGR and 1S:GFP by transmission EM. True leaves from 10-day-old Arabidopsis WT seedlings were processed by HPF and embedded with Lowicryl HM20. ( h – j ) 1S:GFP was detected with Ab-5450 and anti-goat-IgG (18 nm particle). ( k ) Negative control without Ab-CD1-i and Ab-5450. ( l ) Double immunolocalization of 1S:GFP (18 nm particle) and HMGR (12 nm particle). ( m ) HMGR was detected with Ab-CD1-i and anti-rabbit-IgG (12 nm particle). Chloroplast (Chl). ER strands (red arrowheads). Golgi apparatus (G). Mitochondria (M). Nuclear envelope (blue arrowheads). Nucleus (N). Bars, 500 nm.
Alexa Fluor 594, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 99/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/alexa fluor 594/product/Jackson Immuno
Average 99 stars, based on 40 article reviews
Price from $9.99 to $1999.99
alexa fluor 594 - by Bioz Stars, 2022-10
99/100 stars

Images

1) Product Images from "Loose Morphology and High Dynamism of OSER Structures Induced by the Membrane Domain of HMG-CoA Reductase"

Article Title: Loose Morphology and High Dynamism of OSER Structures Induced by the Membrane Domain of HMG-CoA Reductase

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms22179132

Arabidopsis HMGR localizes in the ER network, nuclear envelope, HMGR vesicles and ER-HMGR domains. ( a – c ) Whole-mount immunohistochemical analysis of cotyledon parenchymal cells from 6-day-old Arabidopsis WT seedlings ( a ) Immunodetection of HMGR with Ab-CD1-i and anti-rabbit IgG secondary antibody (Alexa Fluor 555, in red), visualized by confocal microscopy under dark (left) or bright fields (right). ( b ) Immunodetection of HMGR with Ab-CD1-i and anti-rabbit IgG secondary antibody (AlexaFluor 555, in red), and simultaneous detection of chlorophyll (in blue). ( c ) Negative control without the Ab-CD1-i antibody. The irregular corpuscles, 0.2 to 2 µm in length, correspond to HMGR vesicles. The elliptic bodies, 6 to 8 µm in diameter, correspond to chloroplasts. Images were obtained by Z-projection encompassing 3 ( a ), 10 ( b ) or 4 ( c ) µm in the Z-axis. Bars, 5 µm. ( d – f ) Immunochemical study of HMGR vesicles by transmission EM. Leaf samples from 10-day-old Arabidopsis WT or 1S:GFP seedlings were processed by HPF and embedded with Lowicryl HM20. ( d ) HMGR was detected in cotyledon from WT seedlings with Ab-CD1-i and anti-rabbit-IgG (18 nm particle). ( e ) HMGR was detected in true leaf from 1S:GFP seedlings with Ab-CD1-i and anti-rabbit-IgG (12 nm particle). ( f ) Double immunolocalization of HMGR and 1S:GFP in true leaf from 1S:GFP seedlings. HMGR was detected with Ab-CD1-i and anti-rabbit-IgG (12 nm particle) and 1S:GFP was detected with Ab-5450 and anti-goat-IgG (18 nm particle). Black and blue arrowheads indicate, respectively, the external and internal membranes from HMGR vesicles. Red arrowheads indicate ER strands. Bars, 250 nm. ( g ) Whole-mount immunohistochemical analysis of cotyledon parenchymal cells from 6-day-old Arabidopsis 1S:GFP transgenic seedlings. 1S:GFP was detected with Ab-5450 and secondary antibody Alexa fluor 488 (green). HMGR was detected with Ab-CD1-i and secondary antibody Alexa fluor 594 (red). Images were obtained by Z-projection encompassing 12 µm in the Z-axis. Bar, 5 µm. ( h – m ) Immunolocalization of HMGR and 1S:GFP by transmission EM. True leaves from 10-day-old Arabidopsis WT seedlings were processed by HPF and embedded with Lowicryl HM20. ( h – j ) 1S:GFP was detected with Ab-5450 and anti-goat-IgG (18 nm particle). ( k ) Negative control without Ab-CD1-i and Ab-5450. ( l ) Double immunolocalization of 1S:GFP (18 nm particle) and HMGR (12 nm particle). ( m ) HMGR was detected with Ab-CD1-i and anti-rabbit-IgG (12 nm particle). Chloroplast (Chl). ER strands (red arrowheads). Golgi apparatus (G). Mitochondria (M). Nuclear envelope (blue arrowheads). Nucleus (N). Bars, 500 nm.
Figure Legend Snippet: Arabidopsis HMGR localizes in the ER network, nuclear envelope, HMGR vesicles and ER-HMGR domains. ( a – c ) Whole-mount immunohistochemical analysis of cotyledon parenchymal cells from 6-day-old Arabidopsis WT seedlings ( a ) Immunodetection of HMGR with Ab-CD1-i and anti-rabbit IgG secondary antibody (Alexa Fluor 555, in red), visualized by confocal microscopy under dark (left) or bright fields (right). ( b ) Immunodetection of HMGR with Ab-CD1-i and anti-rabbit IgG secondary antibody (AlexaFluor 555, in red), and simultaneous detection of chlorophyll (in blue). ( c ) Negative control without the Ab-CD1-i antibody. The irregular corpuscles, 0.2 to 2 µm in length, correspond to HMGR vesicles. The elliptic bodies, 6 to 8 µm in diameter, correspond to chloroplasts. Images were obtained by Z-projection encompassing 3 ( a ), 10 ( b ) or 4 ( c ) µm in the Z-axis. Bars, 5 µm. ( d – f ) Immunochemical study of HMGR vesicles by transmission EM. Leaf samples from 10-day-old Arabidopsis WT or 1S:GFP seedlings were processed by HPF and embedded with Lowicryl HM20. ( d ) HMGR was detected in cotyledon from WT seedlings with Ab-CD1-i and anti-rabbit-IgG (18 nm particle). ( e ) HMGR was detected in true leaf from 1S:GFP seedlings with Ab-CD1-i and anti-rabbit-IgG (12 nm particle). ( f ) Double immunolocalization of HMGR and 1S:GFP in true leaf from 1S:GFP seedlings. HMGR was detected with Ab-CD1-i and anti-rabbit-IgG (12 nm particle) and 1S:GFP was detected with Ab-5450 and anti-goat-IgG (18 nm particle). Black and blue arrowheads indicate, respectively, the external and internal membranes from HMGR vesicles. Red arrowheads indicate ER strands. Bars, 250 nm. ( g ) Whole-mount immunohistochemical analysis of cotyledon parenchymal cells from 6-day-old Arabidopsis 1S:GFP transgenic seedlings. 1S:GFP was detected with Ab-5450 and secondary antibody Alexa fluor 488 (green). HMGR was detected with Ab-CD1-i and secondary antibody Alexa fluor 594 (red). Images were obtained by Z-projection encompassing 12 µm in the Z-axis. Bar, 5 µm. ( h – m ) Immunolocalization of HMGR and 1S:GFP by transmission EM. True leaves from 10-day-old Arabidopsis WT seedlings were processed by HPF and embedded with Lowicryl HM20. ( h – j ) 1S:GFP was detected with Ab-5450 and anti-goat-IgG (18 nm particle). ( k ) Negative control without Ab-CD1-i and Ab-5450. ( l ) Double immunolocalization of 1S:GFP (18 nm particle) and HMGR (12 nm particle). ( m ) HMGR was detected with Ab-CD1-i and anti-rabbit-IgG (12 nm particle). Chloroplast (Chl). ER strands (red arrowheads). Golgi apparatus (G). Mitochondria (M). Nuclear envelope (blue arrowheads). Nucleus (N). Bars, 500 nm.

Techniques Used: Immunohistochemistry, Immunodetection, Confocal Microscopy, Negative Control, Transmission Assay, Transgenic Assay

2) Product Images from "In vivo neuronal and astrocytic activation in somatosensory cortex by acupuncture stimuli"

Article Title: In vivo neuronal and astrocytic activation in somatosensory cortex by acupuncture stimuli

Journal: Neural Regeneration Research

doi: 10.4103/1673-5374.339003

Acupuncture activates calcium transients in astrocytes . (A) Transfection sites of AAV-gfaABC1D-GCaMP6f in S1 were colocalized with astrocyte marker S100β (white arrowheads, red, stained by Alexa Fluor 594). Scale bars: 50 μm. (B) Pseudo-colored images for the intensity of astrocytic calcium spikes in selected fields of view during the resting state and NS. Scale bars: 50 μm. (C–F) Time-series recordings of normalized calcium values (in ΔF/F 0 ) during the acupuncture sessions (grey shaded box). The temporal scale (x-axis) was presented in seconds. The protocol of acupuncture was the same as in Figure 2 . In each group, the calcium activity from different fields of view was recorded for data plotting. Field of view numbers: n = 9 for C–D, n = 5 for E, n = 10 for F. Animal numbers: n = 3 each. (G) Comparison of total integrated astrocytic calcium activity. Data are expressed as mean ± SEM and were analyzed by one-way analysis of variance. Group effect: F (3, 29) = 5.188, P = 0.0054; Bonferroni’s multiple comparison test: * P
Figure Legend Snippet: Acupuncture activates calcium transients in astrocytes . (A) Transfection sites of AAV-gfaABC1D-GCaMP6f in S1 were colocalized with astrocyte marker S100β (white arrowheads, red, stained by Alexa Fluor 594). Scale bars: 50 μm. (B) Pseudo-colored images for the intensity of astrocytic calcium spikes in selected fields of view during the resting state and NS. Scale bars: 50 μm. (C–F) Time-series recordings of normalized calcium values (in ΔF/F 0 ) during the acupuncture sessions (grey shaded box). The temporal scale (x-axis) was presented in seconds. The protocol of acupuncture was the same as in Figure 2 . In each group, the calcium activity from different fields of view was recorded for data plotting. Field of view numbers: n = 9 for C–D, n = 5 for E, n = 10 for F. Animal numbers: n = 3 each. (G) Comparison of total integrated astrocytic calcium activity. Data are expressed as mean ± SEM and were analyzed by one-way analysis of variance. Group effect: F (3, 29) = 5.188, P = 0.0054; Bonferroni’s multiple comparison test: * P

Techniques Used: Transfection, Marker, Staining, Activity Assay

3) Product Images from "An engineered mutant of a host phospholipid synthesis gene inhibits viral replication without compromising host fitness"

Article Title: An engineered mutant of a host phospholipid synthesis gene inhibits viral replication without compromising host fitness

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.RA118.007051

BMV 1a mislocalizes in the presence of Cho2p-aia. A , microscopy images showing the distribution patterns of Cho2p-HA and Cho2p-aia-HA in cho2 Δ cells in the absence ( w/o ) or presence ( w/ ) of BMV 1a-His 6 . Cho2p-HA and Cho2p-aia-HA were detected using an anti-HA pAb and a secondary antibody conjugated to Alexa Fluor 594. BMV 1a was detected using an anti-His 6 mAb and a secondary antibody conjugated to Alexa Fluor 488. B , distribution patterns of BMV 1a-mCherry in cho2 Δ cells in the absence or presence of Cho2p-aia-HA. C , distribution pattern of BMV 1a-mCherry in cho2 Δ cells in the presence of Cho2p-aia-HA and GFP-tagged organelle markers for inclusion bodies (VHL), Golgi (Sed5p), or ER membrane (Scs2p). Nuclei were stained with DAPI, and scale bars are 2.5 μm in all panels.
Figure Legend Snippet: BMV 1a mislocalizes in the presence of Cho2p-aia. A , microscopy images showing the distribution patterns of Cho2p-HA and Cho2p-aia-HA in cho2 Δ cells in the absence ( w/o ) or presence ( w/ ) of BMV 1a-His 6 . Cho2p-HA and Cho2p-aia-HA were detected using an anti-HA pAb and a secondary antibody conjugated to Alexa Fluor 594. BMV 1a was detected using an anti-His 6 mAb and a secondary antibody conjugated to Alexa Fluor 488. B , distribution patterns of BMV 1a-mCherry in cho2 Δ cells in the absence or presence of Cho2p-aia-HA. C , distribution pattern of BMV 1a-mCherry in cho2 Δ cells in the presence of Cho2p-aia-HA and GFP-tagged organelle markers for inclusion bodies (VHL), Golgi (Sed5p), or ER membrane (Scs2p). Nuclei were stained with DAPI, and scale bars are 2.5 μm in all panels.

Techniques Used: Microscopy, Staining

4) Product Images from "Non-destructive and Selective Imaging of the Functionally Active, Pro-invasive Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) Enzyme in Cancer Cells *"

Article Title: Non-destructive and Selective Imaging of the Functionally Active, Pro-invasive Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) Enzyme in Cancer Cells *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.471508

MP-3653 binds MT3-MMP but not MT6-MMP. A , left panels , MCF7-MT3 and MCF7-MT6 cells were stained using the M2-FLAG antibody followed by secondary anti-mouse antibody conjugated with Alexa Fluor 594. Right two panels , cells were co-incubated at 37 °C
Figure Legend Snippet: MP-3653 binds MT3-MMP but not MT6-MMP. A , left panels , MCF7-MT3 and MCF7-MT6 cells were stained using the M2-FLAG antibody followed by secondary anti-mouse antibody conjugated with Alexa Fluor 594. Right two panels , cells were co-incubated at 37 °C

Techniques Used: Staining, Incubation

MP-3653 binds murine MT1-MMP. A , B16F1-mock and B16F1-mMT1 cells were stained using the MT1-MMP 3G4 antibody followed by the secondary anti-mouse antibody conjugated with Alexa Fluor 594. Right two panels , B16F1-mock and B16F1-mMT1 cells were co-incubated
Figure Legend Snippet: MP-3653 binds murine MT1-MMP. A , B16F1-mock and B16F1-mMT1 cells were stained using the MT1-MMP 3G4 antibody followed by the secondary anti-mouse antibody conjugated with Alexa Fluor 594. Right two panels , B16F1-mock and B16F1-mMT1 cells were co-incubated

Techniques Used: Staining, Incubation

The imaging of cells. Left panels , MCF7-mock, MCF7-MT1, and MCF7-ΔCT cells were stained using the MT1-MMP 3G4 antibody, followed by the secondary anti-mouse antibody conjugated with Alexa Fluor 594. MT1-MMP ( red ), DAPI ( blue ). Right three panels
Figure Legend Snippet: The imaging of cells. Left panels , MCF7-mock, MCF7-MT1, and MCF7-ΔCT cells were stained using the MT1-MMP 3G4 antibody, followed by the secondary anti-mouse antibody conjugated with Alexa Fluor 594. MT1-MMP ( red ), DAPI ( blue ). Right three panels

Techniques Used: Imaging, Staining

5) Product Images from "AFAP-110 is overexpressed in prostate cancer and contributes to tumorigenic growth by regulating focal contacts"

Article Title: AFAP-110 is overexpressed in prostate cancer and contributes to tumorigenic growth by regulating focal contacts

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI30710

Effects of AFAP-110 downregulation on focal adhesion and integrin β1 expression. ( A ) Loss of focal adhesion structures (white arrows) mediated by AFAP-110 downregulation. Immunofluorescence staining of vinculin, a cytoskeletal protein localized at focal adhesion structures, was performed using a monoclonal anti-vinculin primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Actin was stained by Alexa Fluor 488–conjugated phalloidin (green). Representative images of 2 independent experiments are shown; scale bars: 20 μm. ( B ) Expression and activity of Src in phosphorylating adhesion-associated substrates. Western blotting of cell lysates was performed with monoclonal mouse antibodies selective for phospho-FAK tyrosine 861 (Y861), phospho–c-Src tyrosine 418 (Y418), phospho-paxillin tyrosine 118 (Y118), as well as antibodies recognizing total protein Src, FAK, paxillin, and vinculin. Cell lysate from PC3 cells that express a constitutively active form of Src (PC3 Src 527F) was used as a positive control. Membranes were probed for β-actin as loading control. Results from 1 of 2 independent experiments are shown. ( C ) Immunoblotting with antibodies against AFAP-110, integrin β1, and E-cadherin. β-Actin expression was used as a loading control. ( D ) Immunofluorescence staining of integrin β1 was performed as described in Methods, using a monoclonal anti–integrin β1 primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Images were converted to grayscale pictures for best visualization of the localization of integrin β1 at focal adhesion structures in PC3 and scrambled control cells (yellow arrows). Representative images of 2 independent experiments are shown; scale bars: 20 μm.
Figure Legend Snippet: Effects of AFAP-110 downregulation on focal adhesion and integrin β1 expression. ( A ) Loss of focal adhesion structures (white arrows) mediated by AFAP-110 downregulation. Immunofluorescence staining of vinculin, a cytoskeletal protein localized at focal adhesion structures, was performed using a monoclonal anti-vinculin primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Actin was stained by Alexa Fluor 488–conjugated phalloidin (green). Representative images of 2 independent experiments are shown; scale bars: 20 μm. ( B ) Expression and activity of Src in phosphorylating adhesion-associated substrates. Western blotting of cell lysates was performed with monoclonal mouse antibodies selective for phospho-FAK tyrosine 861 (Y861), phospho–c-Src tyrosine 418 (Y418), phospho-paxillin tyrosine 118 (Y118), as well as antibodies recognizing total protein Src, FAK, paxillin, and vinculin. Cell lysate from PC3 cells that express a constitutively active form of Src (PC3 Src 527F) was used as a positive control. Membranes were probed for β-actin as loading control. Results from 1 of 2 independent experiments are shown. ( C ) Immunoblotting with antibodies against AFAP-110, integrin β1, and E-cadherin. β-Actin expression was used as a loading control. ( D ) Immunofluorescence staining of integrin β1 was performed as described in Methods, using a monoclonal anti–integrin β1 primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Images were converted to grayscale pictures for best visualization of the localization of integrin β1 at focal adhesion structures in PC3 and scrambled control cells (yellow arrows). Representative images of 2 independent experiments are shown; scale bars: 20 μm.

Techniques Used: Expressing, Immunofluorescence, Staining, Activity Assay, Western Blot, Positive Control

Restoration of integrin β1 expression and focal adhesions by ectopic expression of wild-type AFAP-110 and functional mutant variants. ( A ) Schematic representation of wild-type AFAP-110 and functional mutant variants. Deletion of the PH1 domain (AFAP-110 Δ180-226) abolishes the association of AFAP-110 with PKC. A single amino acid mutation that changed a proline residue to an alanine (AFAP71A) at the SH3-binding motif abrogates the ability of AFAP-110 to interact with Src. ( B ) Immunoblotting with antibodies against AFAP-110 and integrin β1. Ectopic expression of GFP-tagged chicken wild-type AFAP-110 and mutant variants was visualized as bands that localized slightly higher than endogenous AFAP-110 on the membrane. Vinculin expression was used as a loading control. ( C ) Immunofluorescence staining of vinculin was performed using a monoclonal anti-vinculin primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Images were converted to grayscale for best visualization of focal adhesion structures (yellow arrows). Ectopic expression of GFP-tagged chicken wild-type AFAP-110 and mutant variants in AFAP-110–downregulated clone 309 were visualized as proteins emitting green fluorescence under microscope (green). Representative images are shown; scale bars: 20 μm.
Figure Legend Snippet: Restoration of integrin β1 expression and focal adhesions by ectopic expression of wild-type AFAP-110 and functional mutant variants. ( A ) Schematic representation of wild-type AFAP-110 and functional mutant variants. Deletion of the PH1 domain (AFAP-110 Δ180-226) abolishes the association of AFAP-110 with PKC. A single amino acid mutation that changed a proline residue to an alanine (AFAP71A) at the SH3-binding motif abrogates the ability of AFAP-110 to interact with Src. ( B ) Immunoblotting with antibodies against AFAP-110 and integrin β1. Ectopic expression of GFP-tagged chicken wild-type AFAP-110 and mutant variants was visualized as bands that localized slightly higher than endogenous AFAP-110 on the membrane. Vinculin expression was used as a loading control. ( C ) Immunofluorescence staining of vinculin was performed using a monoclonal anti-vinculin primary antibody and a goat anti-mouse Alexa Fluor 594–conjugated secondary antibody (red). Images were converted to grayscale for best visualization of focal adhesion structures (yellow arrows). Ectopic expression of GFP-tagged chicken wild-type AFAP-110 and mutant variants in AFAP-110–downregulated clone 309 were visualized as proteins emitting green fluorescence under microscope (green). Representative images are shown; scale bars: 20 μm.

Techniques Used: Expressing, Functional Assay, Mutagenesis, Binding Assay, Immunofluorescence, Staining, Fluorescence, Microscopy

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Jackson Immuno alexa fluor 594 affinipure fab fragment donkey anti mouse igg
    Conception and development of heat-stable <t>antibodies</t> via chemical engineering for high-temperature deep immunostaining. a , Schematic diagram of <t>antibody</t> <t>(Ab)</t> diffusion to reach the deep tissue antigen (Ag) target. K a ( T ) is the association constant of the Ab–Ag binding reactions at a given temperature T , and D eff is the effective diffusion coefficient of free Ab as a function of the antibody spatial location ( r ) and T . ‡ denotes transition state. b , The general relationship between D eff , K a , the percentage of active <t>Abs</t> and T . In a hypothetical heat-facilitated strategy, the Ab–Ag binding reaction is not favored at higher T (that is, it lowers K a ), but is also irreversibly denatured at sufficiently high T (brown solid line). Therefore, raising T to increase the free Ab proportion is viable only if the Abs can be protected from denaturation (brown dotted line). c , Reaction–diffusion simulation of Ab–Ag binding and Ab diffusion in a cylindrical arena. The time ( t )-dependent concentration profiles of the Ab–Ag complex ([Ab–Ag]) along the diffusion distance ( r ) with different combinations of T -dependent Ab–Ag binding kinetics and Ab diffusivity are visualized (lower panels). d , [Ab–Ag] versus r at the end of the simulations in c . e , Strategies for stabilizing Abs against permanent heat denaturation: stage 1, complexation with a secondary <t>Fab</t> <t>fragment</t> to stabilize protein conformation; stage 2, multifunctional crosslinkers are used to crosslink the complex (insets). f , SDS–PAGE analysis on crosslinking primary Ab–Fab fragment complexes. AF594, <t>Alexa</t> Fluor 594; C H , heavy chain constant domain; C L , light chain constant domain; FITC, fluorescein isothiocyanate; MW, molecular weight; V H , heavy chain variable domain; V L , light chain variable domain. g , Gel filtration analysis of the optimized <t>IgG–Fab</t> complex crosslinking reaction mixture, including a pair of IgG and Fab with mismatched Fab host specificity. Gt, goat; Rb, rabbit. Albumin peaks have been removed for clarity. The full traces are shown in Extended Data Fig. 2 .
    Alexa Fluor 594 Affinipure Fab Fragment Donkey Anti Mouse Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa fluor 594 affinipure fab fragment donkey anti mouse igg/product/Jackson Immuno
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa fluor 594 affinipure fab fragment donkey anti mouse igg - by Bioz Stars, 2022-10
    90/100 stars
      Buy from Supplier

    97
    Jackson Immuno goat anti mouse igg alexa fluor 594
    MACS maintains the signals of multiple fluorescent probes. a) Fluorescence images of endogenous EYFP signals (1 mm Thy1 ‐YFP‐H brain slices) before and after MACS clearing compared with other clearing protocols. b) Quantification of fluorescence preservation of EYFP, tdTomato, and EGFP after MACS clearing compared with different methods ( n = 3). c) Quantitative analysis of long‐term fluorescence preservation of EYFP, tdTomato, and EGFP after MACS clearing ( n = 3). d) Bright field and fluorescence images of DiI‐labeled brain slices before and after clearing by each method. e) Ultramicroscopic imaging of mouse brain samples restored from MACS or PBS by transmission electron microscopy. Red arrow heads indicate typical membrane structures. f) Fluorescence signals labeled by various chemical fluorescent tracers are finely imaged after MACS clearing, including DiI, PI, tetramethylrhodamine (rhodamine), <t>Alexa</t> <t>Fluor</t> 594 (AF 594)‐conjugated antibody, DsRed, and mCherry. All values are presented as the mean ± SD. Statistical significance in (b) (***, p
    Goat Anti Mouse Igg Alexa Fluor 594, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse igg alexa fluor 594/product/Jackson Immuno
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg alexa fluor 594 - by Bioz Stars, 2022-10
    97/100 stars
      Buy from Supplier

    88
    Jackson Immuno af594
    Overlay of fluorescence images of <t>AF594</t> (Purified Mab), PE (Commercial Mab), and Hoechst at selected antibody concentrations for purified and commercial antibodies. The exposure times are 400000 and 800000 μs, respectively. The fluorescence intensities were clearly reduced as the antibody concentrations decreased.
    Af594, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/af594/product/Jackson Immuno
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    af594 - by Bioz Stars, 2022-10
    88/100 stars
      Buy from Supplier

    Image Search Results


    Conception and development of heat-stable antibodies via chemical engineering for high-temperature deep immunostaining. a , Schematic diagram of antibody (Ab) diffusion to reach the deep tissue antigen (Ag) target. K a ( T ) is the association constant of the Ab–Ag binding reactions at a given temperature T , and D eff is the effective diffusion coefficient of free Ab as a function of the antibody spatial location ( r ) and T . ‡ denotes transition state. b , The general relationship between D eff , K a , the percentage of active Abs and T . In a hypothetical heat-facilitated strategy, the Ab–Ag binding reaction is not favored at higher T (that is, it lowers K a ), but is also irreversibly denatured at sufficiently high T (brown solid line). Therefore, raising T to increase the free Ab proportion is viable only if the Abs can be protected from denaturation (brown dotted line). c , Reaction–diffusion simulation of Ab–Ag binding and Ab diffusion in a cylindrical arena. The time ( t )-dependent concentration profiles of the Ab–Ag complex ([Ab–Ag]) along the diffusion distance ( r ) with different combinations of T -dependent Ab–Ag binding kinetics and Ab diffusivity are visualized (lower panels). d , [Ab–Ag] versus r at the end of the simulations in c . e , Strategies for stabilizing Abs against permanent heat denaturation: stage 1, complexation with a secondary Fab fragment to stabilize protein conformation; stage 2, multifunctional crosslinkers are used to crosslink the complex (insets). f , SDS–PAGE analysis on crosslinking primary Ab–Fab fragment complexes. AF594, Alexa Fluor 594; C H , heavy chain constant domain; C L , light chain constant domain; FITC, fluorescein isothiocyanate; MW, molecular weight; V H , heavy chain variable domain; V L , light chain variable domain. g , Gel filtration analysis of the optimized IgG–Fab complex crosslinking reaction mixture, including a pair of IgG and Fab with mismatched Fab host specificity. Gt, goat; Rb, rabbit. Albumin peaks have been removed for clarity. The full traces are shown in Extended Data Fig. 2 .

    Journal: Nature Methods

    Article Title: Antibody stabilization for thermally accelerated deep immunostaining

    doi: 10.1038/s41592-022-01569-1

    Figure Lengend Snippet: Conception and development of heat-stable antibodies via chemical engineering for high-temperature deep immunostaining. a , Schematic diagram of antibody (Ab) diffusion to reach the deep tissue antigen (Ag) target. K a ( T ) is the association constant of the Ab–Ag binding reactions at a given temperature T , and D eff is the effective diffusion coefficient of free Ab as a function of the antibody spatial location ( r ) and T . ‡ denotes transition state. b , The general relationship between D eff , K a , the percentage of active Abs and T . In a hypothetical heat-facilitated strategy, the Ab–Ag binding reaction is not favored at higher T (that is, it lowers K a ), but is also irreversibly denatured at sufficiently high T (brown solid line). Therefore, raising T to increase the free Ab proportion is viable only if the Abs can be protected from denaturation (brown dotted line). c , Reaction–diffusion simulation of Ab–Ag binding and Ab diffusion in a cylindrical arena. The time ( t )-dependent concentration profiles of the Ab–Ag complex ([Ab–Ag]) along the diffusion distance ( r ) with different combinations of T -dependent Ab–Ag binding kinetics and Ab diffusivity are visualized (lower panels). d , [Ab–Ag] versus r at the end of the simulations in c . e , Strategies for stabilizing Abs against permanent heat denaturation: stage 1, complexation with a secondary Fab fragment to stabilize protein conformation; stage 2, multifunctional crosslinkers are used to crosslink the complex (insets). f , SDS–PAGE analysis on crosslinking primary Ab–Fab fragment complexes. AF594, Alexa Fluor 594; C H , heavy chain constant domain; C L , light chain constant domain; FITC, fluorescein isothiocyanate; MW, molecular weight; V H , heavy chain variable domain; V L , light chain variable domain. g , Gel filtration analysis of the optimized IgG–Fab complex crosslinking reaction mixture, including a pair of IgG and Fab with mismatched Fab host specificity. Gt, goat; Rb, rabbit. Albumin peaks have been removed for clarity. The full traces are shown in Extended Data Fig. 2 .

    Article Snippet: The Fab fragments of secondary antibodies used were Alexa Fluor 594-conjugated donkey anti-goat IgG Fab fragments (cat. no. 705-547-003, Jackson ImmunoResearch), unconjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-007-003, Jackson ImmunoResearch), Alexa Fluor 488-conjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-547-003, Jackson ImmunoResearch), Alexa Fluor 594-conjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-587-003, Jackson ImmunoResearch), Alexa Fluor 647-conjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-607-003, Jackson ImmunoResearch), unconjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-007-003 Jackson ImmunoResearch), Alexa Fluor 488-conjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-547-003, Jackson ImmunoResearch), Alexa Fluor 594-conjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-587-003, Jackson ImmunoResearch), Alexa Fluor 647-conjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-607-003, Jackson ImmunoResearch), and Alexa Fluor 488-conjugated goat anti-rat IgG Fab fragments (cat. no. 112-547-003, Jackson ImmunoResearch).

    Techniques: Immunostaining, Diffusion-based Assay, Binding Assay, Concentration Assay, SDS Page, Molecular Weight, Filtration

    Development and applications of deep immunostaining using thermostabilized antibody–Fab complex. a , Tolerance of SPEARs to the duration and condition of ThICK staining at 55 °C. Upper panels: x – z view of mouse spinal cords that have been ThICK-stained with ChAT SPEARs. Scale bars, 50 μm. Lower panels, example cells from different depths. Scale bars, 10 μm. Color bar, pixel intensity. b , From left to right: homogeneity of pixel intensity mean, variability (that is, s.d.), and signal to noise ratio (SNR) by depth for various durations of ThICK staining (in hours). c , ThICK staining of formaldehyde-fixed (left panel, 55 °C for 1 h) and SHIELD-protected (right panel, 55 °C for 16 h) tissues with endogenous fluorescence. Scale bars, 50 μm. d , Antibodies compatible with SPEAR synthesis and ThICK staining. Green, Alexa Fluor 488; red, Alexa Fluor 594; cyan, Alexa Fluor 647. MIP, maximum intensity projection; PV, parvalbumin; TH, tyrosine hydroxylase; VIP, vasoactive intestinal peptide. Scale bars, 20 μm. e , ThICK staining with NanoSPEARs. Scale bars, 20 μm. f , Optimization of ThICK-staining buffer composition with respect to SPEARs intravascular precipitates per unit imaged tissue volume. The experiment was repeated two more times for control (0.3% Triton X-100) and 1 M GnCl. The error bars represent s.d. P = 0.0216, two-sided unpaired t -test with Welch’s correction. g , Pyridine (py)-catalyzed P3PE-crosslinking reaction. h , A higher concentration of py is associated with more conversion of precursor to product. i , A total of 61.8 mM py accelerates crosslinking when compared with non-catalyzed control by SDS–PAGE. Inset: results of 1–8 h reaction time. *** P ≤ 0.01 at 4 and 8 h reaction (multiple two-sided t -test with multiple comparisons adjustment), n = 3 replicates; data are given as mean ± s.d. j , Functional assay based on hot-start PCR for testing py-catalyzed synthesized SPEARs (SPEAR py ) and agarose gel analysis of the PCR product in the lower panel. k , Functional activity of Taq SPEAR versus Taq SPEAR py in inhibiting PCR product formation, comparing SPEARs used directly after synthesis with those pre-heated at 55 °C for 16 h. The experiment was repeated six times and data are given as mean ± s.d. n.s., not significant. *** P ≤ 0.001 (Tukey’s multiple comparison test, two-sided). l , Comparison of staining qualities of ChAT SPEAR and ChAT SPEAR py . Left panels, representative images of staining with ChAT SPEARs. Scale bars, 20 μm. Right panel, signal to background ratio along the axes of representative cells (white rectangles). Data are given as mean (solid lines) ± s.d. (shaded regions). Lighter lines represent normalized intensity profiles of individual cells. m , Optimized ThICK staining with ChAT SPEAR py (red) in a SHIELD-protected sample with endogenous neuronal GCaMP6f (green). Precipitates were identified and highlighted in white. Scale bars, 50 μm. Source data

    Journal: Nature Methods

    Article Title: Antibody stabilization for thermally accelerated deep immunostaining

    doi: 10.1038/s41592-022-01569-1

    Figure Lengend Snippet: Development and applications of deep immunostaining using thermostabilized antibody–Fab complex. a , Tolerance of SPEARs to the duration and condition of ThICK staining at 55 °C. Upper panels: x – z view of mouse spinal cords that have been ThICK-stained with ChAT SPEARs. Scale bars, 50 μm. Lower panels, example cells from different depths. Scale bars, 10 μm. Color bar, pixel intensity. b , From left to right: homogeneity of pixel intensity mean, variability (that is, s.d.), and signal to noise ratio (SNR) by depth for various durations of ThICK staining (in hours). c , ThICK staining of formaldehyde-fixed (left panel, 55 °C for 1 h) and SHIELD-protected (right panel, 55 °C for 16 h) tissues with endogenous fluorescence. Scale bars, 50 μm. d , Antibodies compatible with SPEAR synthesis and ThICK staining. Green, Alexa Fluor 488; red, Alexa Fluor 594; cyan, Alexa Fluor 647. MIP, maximum intensity projection; PV, parvalbumin; TH, tyrosine hydroxylase; VIP, vasoactive intestinal peptide. Scale bars, 20 μm. e , ThICK staining with NanoSPEARs. Scale bars, 20 μm. f , Optimization of ThICK-staining buffer composition with respect to SPEARs intravascular precipitates per unit imaged tissue volume. The experiment was repeated two more times for control (0.3% Triton X-100) and 1 M GnCl. The error bars represent s.d. P = 0.0216, two-sided unpaired t -test with Welch’s correction. g , Pyridine (py)-catalyzed P3PE-crosslinking reaction. h , A higher concentration of py is associated with more conversion of precursor to product. i , A total of 61.8 mM py accelerates crosslinking when compared with non-catalyzed control by SDS–PAGE. Inset: results of 1–8 h reaction time. *** P ≤ 0.01 at 4 and 8 h reaction (multiple two-sided t -test with multiple comparisons adjustment), n = 3 replicates; data are given as mean ± s.d. j , Functional assay based on hot-start PCR for testing py-catalyzed synthesized SPEARs (SPEAR py ) and agarose gel analysis of the PCR product in the lower panel. k , Functional activity of Taq SPEAR versus Taq SPEAR py in inhibiting PCR product formation, comparing SPEARs used directly after synthesis with those pre-heated at 55 °C for 16 h. The experiment was repeated six times and data are given as mean ± s.d. n.s., not significant. *** P ≤ 0.001 (Tukey’s multiple comparison test, two-sided). l , Comparison of staining qualities of ChAT SPEAR and ChAT SPEAR py . Left panels, representative images of staining with ChAT SPEARs. Scale bars, 20 μm. Right panel, signal to background ratio along the axes of representative cells (white rectangles). Data are given as mean (solid lines) ± s.d. (shaded regions). Lighter lines represent normalized intensity profiles of individual cells. m , Optimized ThICK staining with ChAT SPEAR py (red) in a SHIELD-protected sample with endogenous neuronal GCaMP6f (green). Precipitates were identified and highlighted in white. Scale bars, 50 μm. Source data

    Article Snippet: The Fab fragments of secondary antibodies used were Alexa Fluor 594-conjugated donkey anti-goat IgG Fab fragments (cat. no. 705-547-003, Jackson ImmunoResearch), unconjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-007-003, Jackson ImmunoResearch), Alexa Fluor 488-conjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-547-003, Jackson ImmunoResearch), Alexa Fluor 594-conjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-587-003, Jackson ImmunoResearch), Alexa Fluor 647-conjugated donkey anti-mouse IgG Fab fragments (cat. no. 715-607-003, Jackson ImmunoResearch), unconjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-007-003 Jackson ImmunoResearch), Alexa Fluor 488-conjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-547-003, Jackson ImmunoResearch), Alexa Fluor 594-conjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-587-003, Jackson ImmunoResearch), Alexa Fluor 647-conjugated donkey anti-rabbit IgG Fab fragments (cat. no. 711-607-003, Jackson ImmunoResearch), and Alexa Fluor 488-conjugated goat anti-rat IgG Fab fragments (cat. no. 112-547-003, Jackson ImmunoResearch).

    Techniques: Immunostaining, Staining, Fluorescence, Concentration Assay, SDS Page, Functional Assay, Hot Start PCR, Synthesized, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Activity Assay

    MACS maintains the signals of multiple fluorescent probes. a) Fluorescence images of endogenous EYFP signals (1 mm Thy1 ‐YFP‐H brain slices) before and after MACS clearing compared with other clearing protocols. b) Quantification of fluorescence preservation of EYFP, tdTomato, and EGFP after MACS clearing compared with different methods ( n = 3). c) Quantitative analysis of long‐term fluorescence preservation of EYFP, tdTomato, and EGFP after MACS clearing ( n = 3). d) Bright field and fluorescence images of DiI‐labeled brain slices before and after clearing by each method. e) Ultramicroscopic imaging of mouse brain samples restored from MACS or PBS by transmission electron microscopy. Red arrow heads indicate typical membrane structures. f) Fluorescence signals labeled by various chemical fluorescent tracers are finely imaged after MACS clearing, including DiI, PI, tetramethylrhodamine (rhodamine), Alexa Fluor 594 (AF 594)‐conjugated antibody, DsRed, and mCherry. All values are presented as the mean ± SD. Statistical significance in (b) (***, p

    Journal: Advanced Science

    Article Title: MACS: Rapid Aqueous Clearing System for 3D Mapping of Intact Organs, MACS: Rapid Aqueous Clearing System for 3D Mapping of Intact Organs

    doi: 10.1002/advs.201903185

    Figure Lengend Snippet: MACS maintains the signals of multiple fluorescent probes. a) Fluorescence images of endogenous EYFP signals (1 mm Thy1 ‐YFP‐H brain slices) before and after MACS clearing compared with other clearing protocols. b) Quantification of fluorescence preservation of EYFP, tdTomato, and EGFP after MACS clearing compared with different methods ( n = 3). c) Quantitative analysis of long‐term fluorescence preservation of EYFP, tdTomato, and EGFP after MACS clearing ( n = 3). d) Bright field and fluorescence images of DiI‐labeled brain slices before and after clearing by each method. e) Ultramicroscopic imaging of mouse brain samples restored from MACS or PBS by transmission electron microscopy. Red arrow heads indicate typical membrane structures. f) Fluorescence signals labeled by various chemical fluorescent tracers are finely imaged after MACS clearing, including DiI, PI, tetramethylrhodamine (rhodamine), Alexa Fluor 594 (AF 594)‐conjugated antibody, DsRed, and mCherry. All values are presented as the mean ± SD. Statistical significance in (b) (***, p

    Article Snippet: Secondary antibodies including Alexa Fluor 594 goat anti‐rabbit IgG (H+L) (1:500 dilution; A‐11037, Life Technologies), Alexa Fluor 633 goat anti‐rabbit IgG (H+L) (1:500 dilution; A‐21070, Life Technologies), and goat anti‐mouse IgG Alexa Fluor 594 (1: 300 dilution; 115‐585‐146, Jackson ImmunoResearch Laboratories, Inc.) were used.

    Techniques: Magnetic Cell Separation, Fluorescence, Preserving, Labeling, Imaging, Transmission Assay, Electron Microscopy

    Overlay of fluorescence images of AF594 (Purified Mab), PE (Commercial Mab), and Hoechst at selected antibody concentrations for purified and commercial antibodies. The exposure times are 400000 and 800000 μs, respectively. The fluorescence intensities were clearly reduced as the antibody concentrations decreased.

    Journal: Journal of immunological methods

    Article Title: Novel high-throughput cell-based hybridoma screening methodology using the Celigo Image Cytometer

    doi: 10.1016/j.jim.2017.04.003

    Figure Lengend Snippet: Overlay of fluorescence images of AF594 (Purified Mab), PE (Commercial Mab), and Hoechst at selected antibody concentrations for purified and commercial antibodies. The exposure times are 400000 and 800000 μs, respectively. The fluorescence intensities were clearly reduced as the antibody concentrations decreased.

    Article Snippet: Cells were then washed with sorting buffer (PBS supplemented with 1% FBS) and stained with the secondary goat anti-mouse IgG (H+L) antibodies labeled with AF594 (Jackson ImmunoResearch) for 30 min at 4°C.

    Techniques: Fluorescence, Purification

    Example fluorescence images and intensity histograms of AF594-CHO at high, medium, low and no binding signals for image and flow cytometers. The optimized binding results validated that all the positive hits from image cytometry were also positive hits on flow cytometry. In addition, wild type CHO cells stained with CFSE did not show AF594 signals, which is indicative of no nonspecific binding.

    Journal: Journal of immunological methods

    Article Title: Novel high-throughput cell-based hybridoma screening methodology using the Celigo Image Cytometer

    doi: 10.1016/j.jim.2017.04.003

    Figure Lengend Snippet: Example fluorescence images and intensity histograms of AF594-CHO at high, medium, low and no binding signals for image and flow cytometers. The optimized binding results validated that all the positive hits from image cytometry were also positive hits on flow cytometry. In addition, wild type CHO cells stained with CFSE did not show AF594 signals, which is indicative of no nonspecific binding.

    Article Snippet: Cells were then washed with sorting buffer (PBS supplemented with 1% FBS) and stained with the secondary goat anti-mouse IgG (H+L) antibodies labeled with AF594 (Jackson ImmunoResearch) for 30 min at 4°C.

    Techniques: Fluorescence, Binding Assay, Flow Cytometry, Cytometry, Staining

    Example fluorescence images and intensity scatter plots for AF594 and CFSE at different dilutions of the primary antibodies. CD39-expressing CHO cell population showed decrease in fluorescence signals as the dilution factor increased.

    Journal: Journal of immunological methods

    Article Title: Novel high-throughput cell-based hybridoma screening methodology using the Celigo Image Cytometer

    doi: 10.1016/j.jim.2017.04.003

    Figure Lengend Snippet: Example fluorescence images and intensity scatter plots for AF594 and CFSE at different dilutions of the primary antibodies. CD39-expressing CHO cell population showed decrease in fluorescence signals as the dilution factor increased.

    Article Snippet: Cells were then washed with sorting buffer (PBS supplemented with 1% FBS) and stained with the secondary goat anti-mouse IgG (H+L) antibodies labeled with AF594 (Jackson ImmunoResearch) for 30 min at 4°C.

    Techniques: Fluorescence, Expressing