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    Name:
    Vector TrueVIEW Autofluorescence Quenching Kit with DAPI
    Description:
    The Vector TrueVIEW Autofluorescence Quenching Kit with DAPI provides a novel way to diminish unwanted autofluorescence from non lipofuscin sources and dramatically improve signal to noise ratio It removes unwanted autofluorescence in tissue sections due to aldehyde fixation red blood cells and structural elements such as collagen and elastin The quenching action of the kit reagents provides a clear unambiguous “true view localization of the target antigen
    Catalog Number:
    SP-8500
    Price:
    None
    Category:
    Protein chemifluorescent detection reagents or kits or substrates
    Size:
    15 ml
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    Structured Review

    Vector Laboratories dapi
    Vector TrueVIEW Autofluorescence Quenching Kit with DAPI
    The Vector TrueVIEW Autofluorescence Quenching Kit with DAPI provides a novel way to diminish unwanted autofluorescence from non lipofuscin sources and dramatically improve signal to noise ratio It removes unwanted autofluorescence in tissue sections due to aldehyde fixation red blood cells and structural elements such as collagen and elastin The quenching action of the kit reagents provides a clear unambiguous “true view localization of the target antigen
    https://www.bioz.com/result/dapi/product/Vector Laboratories
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dapi - by Bioz Stars, 2021-04
    86/100 stars

    Images

    1) Product Images from "Characterization and isolation of highly purified porcine satellite cells"

    Article Title: Characterization and isolation of highly purified porcine satellite cells

    Journal: Cell Death Discovery

    doi: 10.1038/cddiscovery.2017.3

    Pig satellite cells participated in the muscle regeneration in recipient mice and lose the engraftment efficiency after long-term expansion in vitro . ( a ) Four weeks after transplantation, engraftment efficiency of freshly isolated pig satellite cells was determined by Dystrophin and Lamin A/C immunofluorescent staining with muscle sections derived from receipient mice. PBS was injected as negative control. Blue indicated DAPI; red indicated Lamin A/C; green indicated Dystrophin. Scale bars, 20 μ m. ( b ) Lamin A/C, Myh3 immunofluorescent staining with series muscle sections derived from recipient mice 7 days after transplantation. Blue indicated DAPI; red indicated Lamin A/C ( b , left) or Myh3 ( b , right); green indicated Dystrophin. Scale bars, 100 μ m. ( c ) Representative immunofluorescent staining images of Dystrophin and Lamin A/C on mouse sections from mice injected with freshly isolated ( c , left), P2 ( c , middle), and P5 ( c , right) satellite cells. Blue indicated DAPI; red indicated Lamin A/C; green indicated Dystrophin. Scale bars, 100 μ m. ( d ) Quantification of the number of Lamin A/C + fibers in each TA 4 weeks after transplantation. Error bars represented S.E.M. and were based on four independent experiments. Significance was analyzed by student’s t -test. * indicated P
    Figure Legend Snippet: Pig satellite cells participated in the muscle regeneration in recipient mice and lose the engraftment efficiency after long-term expansion in vitro . ( a ) Four weeks after transplantation, engraftment efficiency of freshly isolated pig satellite cells was determined by Dystrophin and Lamin A/C immunofluorescent staining with muscle sections derived from receipient mice. PBS was injected as negative control. Blue indicated DAPI; red indicated Lamin A/C; green indicated Dystrophin. Scale bars, 20 μ m. ( b ) Lamin A/C, Myh3 immunofluorescent staining with series muscle sections derived from recipient mice 7 days after transplantation. Blue indicated DAPI; red indicated Lamin A/C ( b , left) or Myh3 ( b , right); green indicated Dystrophin. Scale bars, 100 μ m. ( c ) Representative immunofluorescent staining images of Dystrophin and Lamin A/C on mouse sections from mice injected with freshly isolated ( c , left), P2 ( c , middle), and P5 ( c , right) satellite cells. Blue indicated DAPI; red indicated Lamin A/C; green indicated Dystrophin. Scale bars, 100 μ m. ( d ) Quantification of the number of Lamin A/C + fibers in each TA 4 weeks after transplantation. Error bars represented S.E.M. and were based on four independent experiments. Significance was analyzed by student’s t -test. * indicated P

    Techniques Used: Mouse Assay, In Vitro, Transplantation Assay, Isolation, Staining, Derivative Assay, Injection, Negative Control

    2) Product Images from "Nuclear removal during terminal lens fiber cell differentiation requires CDK1 activity: appropriating mitosis-related nuclear disassembly"

    Article Title: Nuclear removal during terminal lens fiber cell differentiation requires CDK1 activity: appropriating mitosis-related nuclear disassembly

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.106005

    CDK1 deficiency decreased the phosphorylation of lamin A/C, blocked the entry of DLAD into the nucleus and decreased DNA degradation in maturing lens fiber cells. (A-H,J,K) Primary antibodies to p57 KIP2 (A,D), p27 KIP1 (B,E), pLamin A/C (C,F) and DLAD (H,K) were used on E17.5 Cdk1 L/L (A,B,C,G,H) and MLR10; Cdk1 L/L (D,E,F,J,K) lens sections to detect appropriate antigens. (I,L) TUNEL analysis on E17.5 Cdk1 L/L (I) and MLR10; Cdk1 L/L (L) lens sections revealed DNA degradation. (A,D) p57 KIP2 expression (green stained nuclei) initiated in transitional zone epithelial cells in both control ( Cdk1 L/L ) and Cdk1 -deficient ( MLR10; Cdk1 L/L ) lenses but declined quickly as fiber cells elongated (bracket). (B,E) By contrast, p27 KIP1 persisted in the nuclei of fiber cells deep into the cortex of the control lens (bracket in B) but remained more cortical in the MLR10; Cdk1 L/L lenses (E). (C) Lamin A/C phosphorylation (green foci, dashed circle) initiated near the center of the lens where the organelle-free zone is forming. (H,I) In the same region where pLamin A/C is detected in the control lenses, both DLAD-positive nuclei (lower right inset, H) and TUNEL-positive foci (I, yellow staining, arrows) are found. (F,K,L) MLR10; Cdk1 L/L lenses do not contain pLamin A/C (F), display reduced TUNEL-positive fiber cell foci (L, yellow staining, arrow) and exhibit DLAD accumulation around rather than within late fiber cell nuclei (lower right inset, K). Upper right insets in H and K are high magnifications of cortical fiber cells where DLAD expression is comparatively weak; lower right insets are high magnifications of more mature fiber cells where DLAD expression is clearly evident. Nuclei are counterstained with DAPI, which is blue in A-G,J but pseudocolored red in I and L to enhance the contrast for the TUNEL assay. Scale bars: 100 µm in A-F; 200 µm in G-L; 20 µm in H,K (insets).
    Figure Legend Snippet: CDK1 deficiency decreased the phosphorylation of lamin A/C, blocked the entry of DLAD into the nucleus and decreased DNA degradation in maturing lens fiber cells. (A-H,J,K) Primary antibodies to p57 KIP2 (A,D), p27 KIP1 (B,E), pLamin A/C (C,F) and DLAD (H,K) were used on E17.5 Cdk1 L/L (A,B,C,G,H) and MLR10; Cdk1 L/L (D,E,F,J,K) lens sections to detect appropriate antigens. (I,L) TUNEL analysis on E17.5 Cdk1 L/L (I) and MLR10; Cdk1 L/L (L) lens sections revealed DNA degradation. (A,D) p57 KIP2 expression (green stained nuclei) initiated in transitional zone epithelial cells in both control ( Cdk1 L/L ) and Cdk1 -deficient ( MLR10; Cdk1 L/L ) lenses but declined quickly as fiber cells elongated (bracket). (B,E) By contrast, p27 KIP1 persisted in the nuclei of fiber cells deep into the cortex of the control lens (bracket in B) but remained more cortical in the MLR10; Cdk1 L/L lenses (E). (C) Lamin A/C phosphorylation (green foci, dashed circle) initiated near the center of the lens where the organelle-free zone is forming. (H,I) In the same region where pLamin A/C is detected in the control lenses, both DLAD-positive nuclei (lower right inset, H) and TUNEL-positive foci (I, yellow staining, arrows) are found. (F,K,L) MLR10; Cdk1 L/L lenses do not contain pLamin A/C (F), display reduced TUNEL-positive fiber cell foci (L, yellow staining, arrow) and exhibit DLAD accumulation around rather than within late fiber cell nuclei (lower right inset, K). Upper right insets in H and K are high magnifications of cortical fiber cells where DLAD expression is comparatively weak; lower right insets are high magnifications of more mature fiber cells where DLAD expression is clearly evident. Nuclei are counterstained with DAPI, which is blue in A-G,J but pseudocolored red in I and L to enhance the contrast for the TUNEL assay. Scale bars: 100 µm in A-F; 200 µm in G-L; 20 µm in H,K (insets).

    Techniques Used: TUNEL Assay, Expressing, Staining

    MLR10; Cdk1 L/L lens epithelial cells continue to synthesize DNA but fail to enter mitosis. (A-H) BrdU incorporation and phosphorylated histone H3 (pHH3) immunohistochemistry (green nuclei) were used to determine the proportion of cells in S phase (A-D) and late G2 phase (E-H), respectively. Nuclei were counterstained with DAPI. The proliferation index (S-phase fraction) did not differ significantly between MLR10; Cdk1 L/L and Cdk1 L/L lenses at E12.5 or at E15.5 (compare B with A, solid bars in I) but significantly increased in MLR10; Cdk1 L/L lenses by E17.5 (compare D with C, solid bars in I). Although there were fewer overall BrdU-positive nuclei in the MLR10; Cdk1 L/L lenses, the proportion of total nuclei that were BrdU positive was relatively increased at E17.5. The proportion of pHH3-positive cells levels were significantly higher in Cdk1 -deficient lenses beginning at E15.5 (compare E with F, lightly shaded bars in I) and most remaining epithelial cells in MLR10; Cdk1 L/L lenses stained positive for pHH3 by E17.5 (compare H with G, lightly shaded bars in I). (J,K) Whole lenses from Cdk1 L/L (J) and MLR10; Cdk1 L/L mice (K) were stained with DAPI, and the intact epithelium was visualized by confocal microscopy to visualize the size and density of epithelial nuclei. (K,L) MLR10; Cdk1 L/L lenses exhibited a significant increase in nuclear size and an increased DAPI staining foci in each cell. All bars represent the mean ± s.e.m., with each bar representing nine measurements (three sections from each of three different embryos). Scale bars: 100 µm in A-D; 200 µm in E-H; 20 µm J,K. le, lens; re, retina; epi, lens epithelium.
    Figure Legend Snippet: MLR10; Cdk1 L/L lens epithelial cells continue to synthesize DNA but fail to enter mitosis. (A-H) BrdU incorporation and phosphorylated histone H3 (pHH3) immunohistochemistry (green nuclei) were used to determine the proportion of cells in S phase (A-D) and late G2 phase (E-H), respectively. Nuclei were counterstained with DAPI. The proliferation index (S-phase fraction) did not differ significantly between MLR10; Cdk1 L/L and Cdk1 L/L lenses at E12.5 or at E15.5 (compare B with A, solid bars in I) but significantly increased in MLR10; Cdk1 L/L lenses by E17.5 (compare D with C, solid bars in I). Although there were fewer overall BrdU-positive nuclei in the MLR10; Cdk1 L/L lenses, the proportion of total nuclei that were BrdU positive was relatively increased at E17.5. The proportion of pHH3-positive cells levels were significantly higher in Cdk1 -deficient lenses beginning at E15.5 (compare E with F, lightly shaded bars in I) and most remaining epithelial cells in MLR10; Cdk1 L/L lenses stained positive for pHH3 by E17.5 (compare H with G, lightly shaded bars in I). (J,K) Whole lenses from Cdk1 L/L (J) and MLR10; Cdk1 L/L mice (K) were stained with DAPI, and the intact epithelium was visualized by confocal microscopy to visualize the size and density of epithelial nuclei. (K,L) MLR10; Cdk1 L/L lenses exhibited a significant increase in nuclear size and an increased DAPI staining foci in each cell. All bars represent the mean ± s.e.m., with each bar representing nine measurements (three sections from each of three different embryos). Scale bars: 100 µm in A-D; 200 µm in E-H; 20 µm J,K. le, lens; re, retina; epi, lens epithelium.

    Techniques Used: BrdU Incorporation Assay, Immunohistochemistry, Staining, Mouse Assay, Confocal Microscopy

    MLR10; Cdk1 L/L lenses remove both mitochondria and endoplasmic reticulum, despite retaining nuclei. Mitochondria were detected by Tom20 (A,B,D,E) and endoplasmic reticulum by PDI (C,F) immunofluorescence (green staining) in E17.5 lenses. Cdk1 L/L (A-C) and MLR10; Cdk1 L/L (D-F) lens fiber cells lose their mitochondria (A,B,D,E) and endoplasmic reticulum (C,F) prior to reaching the center of the lens. Tom20 staining drops precipitously outside the most peripheral fiber cells (asterisks in A,D) but remains as punctate foci until the deep fiber cells of the central zone (arrows in A,D). Cdk1 L/L lenses form an organelle-free zone (OFZ) lacking both mitochondria and nuclei (B). The MLR10; Cdk1 L/L central lens fibers lack mitochondria, but retain nuclei (E, yellow arrowheads). Likewise, both control (C) and MLR10; Cdk1 L/L (F) lenses remove PDI-staining endoplasmic reticulum from mature nuclear fiber cells (lack of green staining within the dotted line border in C,F). Nuclei were counterstained with DAPI (B,E). Scale bars: 100 µm A,B,D,E; 200 µm C,F. epi, lens epithelium; cf, central fiber cells.
    Figure Legend Snippet: MLR10; Cdk1 L/L lenses remove both mitochondria and endoplasmic reticulum, despite retaining nuclei. Mitochondria were detected by Tom20 (A,B,D,E) and endoplasmic reticulum by PDI (C,F) immunofluorescence (green staining) in E17.5 lenses. Cdk1 L/L (A-C) and MLR10; Cdk1 L/L (D-F) lens fiber cells lose their mitochondria (A,B,D,E) and endoplasmic reticulum (C,F) prior to reaching the center of the lens. Tom20 staining drops precipitously outside the most peripheral fiber cells (asterisks in A,D) but remains as punctate foci until the deep fiber cells of the central zone (arrows in A,D). Cdk1 L/L lenses form an organelle-free zone (OFZ) lacking both mitochondria and nuclei (B). The MLR10; Cdk1 L/L central lens fibers lack mitochondria, but retain nuclei (E, yellow arrowheads). Likewise, both control (C) and MLR10; Cdk1 L/L (F) lenses remove PDI-staining endoplasmic reticulum from mature nuclear fiber cells (lack of green staining within the dotted line border in C,F). Nuclei were counterstained with DAPI (B,E). Scale bars: 100 µm A,B,D,E; 200 µm C,F. epi, lens epithelium; cf, central fiber cells.

    Techniques Used: Immunofluorescence, Staining

    Related Articles

    Staining:

    Article Title: Role of epithelial integrin-linked kinase in promoting intestinal inflammation: effects on CCL2, fibronectin and the T cell repertoire
    Article Snippet: For detection of Th17 cells by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: DAPI and IL-17A (eBio 17B7). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). ..

    Article Title: Uncontrolled angiogenic precursor expansion causes coronary artery anomalies in mice lacking Pofut1
    Article Snippet: After a 2-hour pulse, the hearts were collected and processed for frozen sections as described above in Immunostaining Section. .. The serial sections crossing the whole heart were first stained with PECAM1 or VEGFR3 antibody followed by EdU staining with EdU imaging Kit (Life Technology) and counterstained with DAPI (Vector lab). .. The stained sections were photographed using a Zeiss Observer Z1 or Leica SP5 confocal microscope.

    Article Title: Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter rodentium-induced colitis
    Article Snippet: For detection of Tir and LPS by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: Tir and LPS (Vallance). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). .. A Zeiss LSM-780 microscope was used to capture images.

    Article Title: The EJC component Magoh regulates proliferation and expansion of neural crest-derived melanocytes
    Article Snippet: The following antibodies were used at 1:200: mouse anti- -galactosidase (Promega, madison, WI), rabbit anti-cleaved Caspase-3 (Cell Signaling, Danvers, MA), rabbit anti-phospho-Histone H3 (Upstate Biotechnology, Temecula, CA), and anti-mouse and anti-rabbit FITC-conjugated and rhodamine-conjugated secondary antibodies (Invitrogen, CA). .. Staining was performed using the MOM kit and Vectashield mount with DAPI (Vector labs). ..

    Article Title: Nanoparticle engineered TRAIL-overexpressing adipose-derived stem cells target and eradicate glioblastoma via intracranial delivery
    Article Snippet: Another seven GBM orthotopic mice underwent repetitive injection of NPs/pT RAIL-hADSCs at day 5 and day 15 after inoculation. .. To detect in vivo apoptotic activity, tumor-bearing brain sections were stained with a TUNEL assay kit (Roche) and mounted with Vectashield mounting medium with DAPI (Vector Lab). .. Imaging was performed with the fluorescence microscope (Carl Zeiss).

    Article Title: Preclinical Evaluation of Intravesical Cisplatin Nanoparticles for Non–Muscle-Invasive Bladder Cancer
    Article Snippet: .. Immunofluorescence was performed on a separate set of slides which were stained for TUNEL per manufacturer instructions (ApopTag Peroxidase In Situ Apoptosis Detection Kit, Millipore) and counterstained with DAPI (Vector Laboratories). ..

    Article Title: Platelet-Derived Growth Factor BB Induces Nuclear Export and Proteasomal Degradation of CREB via Phosphatidylinositol 3-Kinase/Akt Signaling in Pulmonary Artery Smooth Muscle Cells
    Article Snippet: SuperSignal West Pico chemiluminescent substrate and NE-PER nuclear and cytoplasmic extract reagents were from Pierce (Rockford, IL). .. Vector NovaRed peroxidase staining kits, VectaShield mounting medium with DAPI (4′,6′-diamidino-2-phenylindole), and peroxidase-conjugated goat anti-rabbit secondary antibody were from Vector Laboratories (Burlingame, CA). .. The plasmid pECFP-C1 was purchased from Clontech Laboratories, Inc. (Palo Alto, CA).

    Conjugation Assay:

    Article Title: Role of epithelial integrin-linked kinase in promoting intestinal inflammation: effects on CCL2, fibronectin and the T cell repertoire
    Article Snippet: For detection of Th17 cells by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: DAPI and IL-17A (eBio 17B7). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). ..

    Article Title: Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter rodentium-induced colitis
    Article Snippet: For detection of Tir and LPS by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: Tir and LPS (Vallance). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). .. A Zeiss LSM-780 microscope was used to capture images.

    Avidin-Biotin Assay:

    Article Title: Role of epithelial integrin-linked kinase in promoting intestinal inflammation: effects on CCL2, fibronectin and the T cell repertoire
    Article Snippet: For detection of Th17 cells by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: DAPI and IL-17A (eBio 17B7). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). ..

    Article Title: Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter rodentium-induced colitis
    Article Snippet: For detection of Tir and LPS by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: Tir and LPS (Vallance). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). .. A Zeiss LSM-780 microscope was used to capture images.

    Immunofluorescence:

    Article Title: Role of epithelial integrin-linked kinase in promoting intestinal inflammation: effects on CCL2, fibronectin and the T cell repertoire
    Article Snippet: For detection of Th17 cells by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: DAPI and IL-17A (eBio 17B7). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). ..

    Article Title: Requirement of epithelial integrin-linked kinase for facilitation of Citrobacter rodentium-induced colitis
    Article Snippet: For detection of Tir and LPS by immunofluorescence, the slides were processed as for IHC and the following antibodies were used: Tir and LPS (Vallance). .. Sections were stained with Vectastain ABC elite kit and biotinylated ant-rabbit for DAPI, or eFluor650 Nanocrystal conjugation kit, cat no. 88-7072-98 antibody, and Avidin D-FITC (or Avidin-Texas Red) used for immunofluorescence (Vector laboratories, CA, USA). .. A Zeiss LSM-780 microscope was used to capture images.

    Article Title: Preclinical Evaluation of Intravesical Cisplatin Nanoparticles for Non–Muscle-Invasive Bladder Cancer
    Article Snippet: .. Immunofluorescence was performed on a separate set of slides which were stained for TUNEL per manufacturer instructions (ApopTag Peroxidase In Situ Apoptosis Detection Kit, Millipore) and counterstained with DAPI (Vector Laboratories). ..

    Microscopy:

    Article Title: Targeting β-tubulin:CCT-β complexes incurs Hsp90- and VCP-related protein degradation and induces ER stress-associated apoptosis by triggering capacitative Ca2+ entry, mitochondrial perturbation and caspase overactivation
    Article Snippet: For mitochondrial staining, cells were incubated with MitoTracker (Invitrogen) for another 15 min at RT. .. After mounting the cells with a commercial kit containing DAPI reagent (Vector Laboratories), the cells were analyzed using a FluoView confocal microscope system (Olympus). .. Determination of caspase activity The caspase activity assay was performed according to the manufacturer's guidelines (BioVision).

    Imaging:

    Article Title: Uncontrolled angiogenic precursor expansion causes coronary artery anomalies in mice lacking Pofut1
    Article Snippet: After a 2-hour pulse, the hearts were collected and processed for frozen sections as described above in Immunostaining Section. .. The serial sections crossing the whole heart were first stained with PECAM1 or VEGFR3 antibody followed by EdU staining with EdU imaging Kit (Life Technology) and counterstained with DAPI (Vector lab). .. The stained sections were photographed using a Zeiss Observer Z1 or Leica SP5 confocal microscope.

    In Vivo:

    Article Title: Nanoparticle engineered TRAIL-overexpressing adipose-derived stem cells target and eradicate glioblastoma via intracranial delivery
    Article Snippet: Another seven GBM orthotopic mice underwent repetitive injection of NPs/pT RAIL-hADSCs at day 5 and day 15 after inoculation. .. To detect in vivo apoptotic activity, tumor-bearing brain sections were stained with a TUNEL assay kit (Roche) and mounted with Vectashield mounting medium with DAPI (Vector Lab). .. Imaging was performed with the fluorescence microscope (Carl Zeiss).

    Activity Assay:

    Article Title: Nanoparticle engineered TRAIL-overexpressing adipose-derived stem cells target and eradicate glioblastoma via intracranial delivery
    Article Snippet: Another seven GBM orthotopic mice underwent repetitive injection of NPs/pT RAIL-hADSCs at day 5 and day 15 after inoculation. .. To detect in vivo apoptotic activity, tumor-bearing brain sections were stained with a TUNEL assay kit (Roche) and mounted with Vectashield mounting medium with DAPI (Vector Lab). .. Imaging was performed with the fluorescence microscope (Carl Zeiss).

    TUNEL Assay:

    Article Title: Nanoparticle engineered TRAIL-overexpressing adipose-derived stem cells target and eradicate glioblastoma via intracranial delivery
    Article Snippet: Another seven GBM orthotopic mice underwent repetitive injection of NPs/pT RAIL-hADSCs at day 5 and day 15 after inoculation. .. To detect in vivo apoptotic activity, tumor-bearing brain sections were stained with a TUNEL assay kit (Roche) and mounted with Vectashield mounting medium with DAPI (Vector Lab). .. Imaging was performed with the fluorescence microscope (Carl Zeiss).

    Article Title: Preclinical Evaluation of Intravesical Cisplatin Nanoparticles for Non–Muscle-Invasive Bladder Cancer
    Article Snippet: .. Immunofluorescence was performed on a separate set of slides which were stained for TUNEL per manufacturer instructions (ApopTag Peroxidase In Situ Apoptosis Detection Kit, Millipore) and counterstained with DAPI (Vector Laboratories). ..

    In Situ:

    Article Title: Preclinical Evaluation of Intravesical Cisplatin Nanoparticles for Non–Muscle-Invasive Bladder Cancer
    Article Snippet: .. Immunofluorescence was performed on a separate set of slides which were stained for TUNEL per manufacturer instructions (ApopTag Peroxidase In Situ Apoptosis Detection Kit, Millipore) and counterstained with DAPI (Vector Laboratories). ..

    Plasmid Preparation:

    Article Title: Platelet-Derived Growth Factor BB Induces Nuclear Export and Proteasomal Degradation of CREB via Phosphatidylinositol 3-Kinase/Akt Signaling in Pulmonary Artery Smooth Muscle Cells
    Article Snippet: SuperSignal West Pico chemiluminescent substrate and NE-PER nuclear and cytoplasmic extract reagents were from Pierce (Rockford, IL). .. Vector NovaRed peroxidase staining kits, VectaShield mounting medium with DAPI (4′,6′-diamidino-2-phenylindole), and peroxidase-conjugated goat anti-rabbit secondary antibody were from Vector Laboratories (Burlingame, CA). .. The plasmid pECFP-C1 was purchased from Clontech Laboratories, Inc. (Palo Alto, CA).

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    Vector Laboratories dapi containing mounting medium
    Peritoneal tumor imaging with ZHS-QDs. Mice bearing peritoneal tumors (PTs) created with MKN45P-luc luciferase-positive human gastric cancer cells received an intraperitoneal co-injection of iRGD or <t>PBS</t> with ZHS-QDs. Intraperitoneal etchant (1x Ag-TS) was given 90 min later. n = 3 per group. Some mice carried asubcutaneous MKN45P-luc tumor (SCT) in addition to the PTs. a , d Whole body in vivo and post-necropsy in situ imaging of the tumor mice, and ex vivo imaging of resected tissues. Lum, luminescence; 800 nm, NIR. The white dotted line in d marks a SCT in a mouse that received iRGD, ZHS-QDs, and Ag-TS. b , e Fluorescent signal per unit area in collected tissues b and in SCT vs. PT in the iRGD group e . c Confocal micrographs of PTs. Blue , <t>DAPI;</t> red , CD31 or ER-TR7; green , ZHS-QDs; scale bars , 50 μm. The white dotted lines indicate PT surface. B, brain; H, heart; Li, liver; S, spleen; Lu, lung; K, kidney. Statistics, two-way analysis of variance b or Student’s t -test e ; error bars , SEM; ns, not significant; ** P
    Dapi Containing Mounting Medium, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories dapi
    Triple AAV Vectors Drive Full-Length Protein Expression in the Mouse Retina (A) Fluorescence analysis of retinal cryosections from C57BL/6J mice 2 months following subretinal injection of triple ED -AAV2/8 vectors under the control of the ubiquitous CMV promoter. The pictures with insets at higher magnification are representative of n = 5 injected eyes. Full arrows indicate EGFP and DsRed double-positive PRs. Dashed arrows indicate EGFP-only positive PRs. The scale bar (20 μm) is depicted in the figure. DsRed, Discosoma red fluorescent protein; Merge, overlay of EGFP, DsRed, and <t>DAPI</t> images; RPE, retinal pigmented epithelium; ONL, outer nuclear layer. (B) Western blot (WB) analysis of truncated products in eyecup lysates from C57BL/6J mice 2 months following subretinal injection of the combinations of ED -AAVs. The arrows indicate the following products: #A, ED full-length protein; #C, from AAV 1 + 3. (C) WB analysis of lysates from C57BL/6J eyecup 2 months following subretinal injection of triple AAV2/8 vectors encoding for ED under the control of the ubiquitous CMV, the RPE-specific VMD2, and the PR-specific IRBP promoters. The arrow indicates the full-length protein. (D) WB analysis of eyecup lysates from C57BL/6J mice 2 months following subretinal injection of single and triple AAV2/8 vectors encoding for ED under the control of the ubiquitous promoter CMV. (E) WB analysis of eyecup lysates from C57BL/6J mice 2 months following subretinal injection of single and triple AAV2/8 vectors encoding for ED under the control of the PR-specific promoter IRBP. CMV, cytomegalovirus; VMD2, vitelliform macular dystrophy 2; IRBP, human interphotoreceptors retinoid binding proteins. α-3xflag, WB with anti-3xflag antibodies; Ponceau, staining with Ponceau, used as loading control; α-β-Tubulin, WB with anti-β-Tubulin antibodies, used as loading control. Neg, lysates of eyecups following injection with <t>PBS.</t> The molecular weight ladder is depicted on the left; 100 (D), 150 (B and C), and 200 (E) μg of proteins were loaded.
    Dapi, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories 6 diamidino 2 phenylindole
    Expression of Rit2 mRNA and protein in the mouse retina. The top panels show immunohistochemistry of retinal flat-mounts ( A - D ) and retinal sections ( E - H ). Images are staining with anti-RIT2 antibody ( A , E ), anti-POU4F2 antibody ( B , F ), merged image of RIT2 and POU4F2 ( C , G ), and nuclear staining by 4', <t>6-diamidino-2-phenylindole</t> (DAPI; D , H ). RIT2-positive cells are more numerous than POU4F2-positive RGCs. E : RIT2 protein is detected in the ganglion cell layer (GCL), inner plexiform layer (IPL), and cells in the inner nuclear layer (INL). Short arrowheads point to RIT2-positive cells located at the inner border of the INL; long arrowheads point to RIT2-positive cells at the outer aspect of the INL. Scale bar: 50 μm. I : This panel shows Rit2 mRNA expression. Laser capture microdissection was used to collect tissues of GCL, INL, and outer nuclear layer (ONL) from mouse retinal sections, and RNA of each layer was analyzed by reverse transcription-quantitative polymerase chain reaction. The mRNA level of Rit2 , Pou4f1 , and Pou4f2 was normalized by that of Gapdh . J : RIT2 protein levels were analyzed at different ages. Protein lysates were prepared from mouse retinas at postnatal day 2 (P2), P5, P15, and adult (8 weeks old), and analyzed by western blotting with anti-RIT2 antibody and anti-α tubulin antibody for control. Protein signals were visualized by chemiluminescence with X-ray films. K : This panel shows relative quantification of RIT2 protein levels. Western blot signals on the X-ray films were scanned and analyzed with ImageJ software. The signal intensity of RIT2 was normalized by that of α tubulin and presented as the RIT2/tubulin ratio. RIT2 protein levels increased from P2 to adults by more than 15 fold.
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    Peritoneal tumor imaging with ZHS-QDs. Mice bearing peritoneal tumors (PTs) created with MKN45P-luc luciferase-positive human gastric cancer cells received an intraperitoneal co-injection of iRGD or PBS with ZHS-QDs. Intraperitoneal etchant (1x Ag-TS) was given 90 min later. n = 3 per group. Some mice carried asubcutaneous MKN45P-luc tumor (SCT) in addition to the PTs. a , d Whole body in vivo and post-necropsy in situ imaging of the tumor mice, and ex vivo imaging of resected tissues. Lum, luminescence; 800 nm, NIR. The white dotted line in d marks a SCT in a mouse that received iRGD, ZHS-QDs, and Ag-TS. b , e Fluorescent signal per unit area in collected tissues b and in SCT vs. PT in the iRGD group e . c Confocal micrographs of PTs. Blue , DAPI; red , CD31 or ER-TR7; green , ZHS-QDs; scale bars , 50 μm. The white dotted lines indicate PT surface. B, brain; H, heart; Li, liver; S, spleen; Lu, lung; K, kidney. Statistics, two-way analysis of variance b or Student’s t -test e ; error bars , SEM; ns, not significant; ** P

    Journal: Nature Communications

    Article Title: In vivo cation exchange in quantum dots for tumor-specific imaging

    doi: 10.1038/s41467-017-00153-y

    Figure Lengend Snippet: Peritoneal tumor imaging with ZHS-QDs. Mice bearing peritoneal tumors (PTs) created with MKN45P-luc luciferase-positive human gastric cancer cells received an intraperitoneal co-injection of iRGD or PBS with ZHS-QDs. Intraperitoneal etchant (1x Ag-TS) was given 90 min later. n = 3 per group. Some mice carried asubcutaneous MKN45P-luc tumor (SCT) in addition to the PTs. a , d Whole body in vivo and post-necropsy in situ imaging of the tumor mice, and ex vivo imaging of resected tissues. Lum, luminescence; 800 nm, NIR. The white dotted line in d marks a SCT in a mouse that received iRGD, ZHS-QDs, and Ag-TS. b , e Fluorescent signal per unit area in collected tissues b and in SCT vs. PT in the iRGD group e . c Confocal micrographs of PTs. Blue , DAPI; red , CD31 or ER-TR7; green , ZHS-QDs; scale bars , 50 μm. The white dotted lines indicate PT surface. B, brain; H, heart; Li, liver; S, spleen; Lu, lung; K, kidney. Statistics, two-way analysis of variance b or Student’s t -test e ; error bars , SEM; ns, not significant; ** P

    Article Snippet: After washing with PBS, sections were mounted in DAPI-containing mounting medium (Vector Laboratories, Burlingame, CA) and examined under a Zeiss LSM 710 NLO confocal microscope.

    Techniques: Imaging, Mouse Assay, Luciferase, Injection, In Vivo, In Situ, Ex Vivo

    Rearrangement of actin cytoskeleton in infected ED cells. ( A ) ED cells were infected with parental, US3_1 -deleted, EHV-1 D207A , US3_4 -recombinant, or revertant viruses and imaged by immunofluorescence microscopy using a Zeiss Axiovert fluorescence microscope. Pictures were taken with a 63x oil objective. The nucleus was stained with 4′,6-diamidino-2-phenylindole (DAPI, blue ), the actin cytoskeleton was stained with phalloidin-Alexa 568 ( red ), and the virus-infected cells were visualized through enhanced GFP (eGFP) expression ( green ). ( B and C ) A total of 200 infected cells for each virus in three independent experiments were inspected and the percentage of infected cells with or without changes in actin cytoskeleton was calculated. A significant change in actin cytoskeleton rearrangement was detected for either EHV-1ΔUS3 or EHV-1 D207A when compared to parental, recombinant, or revertant viruses (one-way ANOVA; p

    Journal: Viruses

    Article Title: The Role of the Equine Herpesvirus Type 1 (EHV-1) US3-Encoded Protein Kinase in Actin Reorganization and Nuclear Egress

    doi: 10.3390/v8100275

    Figure Lengend Snippet: Rearrangement of actin cytoskeleton in infected ED cells. ( A ) ED cells were infected with parental, US3_1 -deleted, EHV-1 D207A , US3_4 -recombinant, or revertant viruses and imaged by immunofluorescence microscopy using a Zeiss Axiovert fluorescence microscope. Pictures were taken with a 63x oil objective. The nucleus was stained with 4′,6-diamidino-2-phenylindole (DAPI, blue ), the actin cytoskeleton was stained with phalloidin-Alexa 568 ( red ), and the virus-infected cells were visualized through enhanced GFP (eGFP) expression ( green ). ( B and C ) A total of 200 infected cells for each virus in three independent experiments were inspected and the percentage of infected cells with or without changes in actin cytoskeleton was calculated. A significant change in actin cytoskeleton rearrangement was detected for either EHV-1ΔUS3 or EHV-1 D207A when compared to parental, recombinant, or revertant viruses (one-way ANOVA; p

    Article Snippet: After blocking with 2% bovine serum albumin (BSA, AppliChem, Berlin, Germany), polymerized actin of infected cells was detected by staining with phalloidin-Alexa-Fluor-568 (1:40; Invitrogen) for 20 min. After washing three times, coverslips were mounted onto glass slides using Vectashield-with 4′,6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Peterborough, UK).

    Techniques: Infection, Recombinant, Immunofluorescence, Microscopy, Fluorescence, Staining, Expressing

    Triple AAV Vectors Drive Full-Length Protein Expression in the Mouse Retina (A) Fluorescence analysis of retinal cryosections from C57BL/6J mice 2 months following subretinal injection of triple ED -AAV2/8 vectors under the control of the ubiquitous CMV promoter. The pictures with insets at higher magnification are representative of n = 5 injected eyes. Full arrows indicate EGFP and DsRed double-positive PRs. Dashed arrows indicate EGFP-only positive PRs. The scale bar (20 μm) is depicted in the figure. DsRed, Discosoma red fluorescent protein; Merge, overlay of EGFP, DsRed, and DAPI images; RPE, retinal pigmented epithelium; ONL, outer nuclear layer. (B) Western blot (WB) analysis of truncated products in eyecup lysates from C57BL/6J mice 2 months following subretinal injection of the combinations of ED -AAVs. The arrows indicate the following products: #A, ED full-length protein; #C, from AAV 1 + 3. (C) WB analysis of lysates from C57BL/6J eyecup 2 months following subretinal injection of triple AAV2/8 vectors encoding for ED under the control of the ubiquitous CMV, the RPE-specific VMD2, and the PR-specific IRBP promoters. The arrow indicates the full-length protein. (D) WB analysis of eyecup lysates from C57BL/6J mice 2 months following subretinal injection of single and triple AAV2/8 vectors encoding for ED under the control of the ubiquitous promoter CMV. (E) WB analysis of eyecup lysates from C57BL/6J mice 2 months following subretinal injection of single and triple AAV2/8 vectors encoding for ED under the control of the PR-specific promoter IRBP. CMV, cytomegalovirus; VMD2, vitelliform macular dystrophy 2; IRBP, human interphotoreceptors retinoid binding proteins. α-3xflag, WB with anti-3xflag antibodies; Ponceau, staining with Ponceau, used as loading control; α-β-Tubulin, WB with anti-β-Tubulin antibodies, used as loading control. Neg, lysates of eyecups following injection with PBS. The molecular weight ladder is depicted on the left; 100 (D), 150 (B and C), and 200 (E) μg of proteins were loaded.

    Journal: Molecular Therapy

    Article Title: Triple Vectors Expand AAV Transfer Capacity in the Retina

    doi: 10.1016/j.ymthe.2017.11.019

    Figure Lengend Snippet: Triple AAV Vectors Drive Full-Length Protein Expression in the Mouse Retina (A) Fluorescence analysis of retinal cryosections from C57BL/6J mice 2 months following subretinal injection of triple ED -AAV2/8 vectors under the control of the ubiquitous CMV promoter. The pictures with insets at higher magnification are representative of n = 5 injected eyes. Full arrows indicate EGFP and DsRed double-positive PRs. Dashed arrows indicate EGFP-only positive PRs. The scale bar (20 μm) is depicted in the figure. DsRed, Discosoma red fluorescent protein; Merge, overlay of EGFP, DsRed, and DAPI images; RPE, retinal pigmented epithelium; ONL, outer nuclear layer. (B) Western blot (WB) analysis of truncated products in eyecup lysates from C57BL/6J mice 2 months following subretinal injection of the combinations of ED -AAVs. The arrows indicate the following products: #A, ED full-length protein; #C, from AAV 1 + 3. (C) WB analysis of lysates from C57BL/6J eyecup 2 months following subretinal injection of triple AAV2/8 vectors encoding for ED under the control of the ubiquitous CMV, the RPE-specific VMD2, and the PR-specific IRBP promoters. The arrow indicates the full-length protein. (D) WB analysis of eyecup lysates from C57BL/6J mice 2 months following subretinal injection of single and triple AAV2/8 vectors encoding for ED under the control of the ubiquitous promoter CMV. (E) WB analysis of eyecup lysates from C57BL/6J mice 2 months following subretinal injection of single and triple AAV2/8 vectors encoding for ED under the control of the PR-specific promoter IRBP. CMV, cytomegalovirus; VMD2, vitelliform macular dystrophy 2; IRBP, human interphotoreceptors retinoid binding proteins. α-3xflag, WB with anti-3xflag antibodies; Ponceau, staining with Ponceau, used as loading control; α-β-Tubulin, WB with anti-β-Tubulin antibodies, used as loading control. Neg, lysates of eyecups following injection with PBS. The molecular weight ladder is depicted on the left; 100 (D), 150 (B and C), and 200 (E) μg of proteins were loaded.

    Article Snippet: Seventy-two hours post-infection, cells were washed once with PBS, fixed for 7 min with 4% paraformaldehyde (PFA) in PBS, washed three times with PBS, and mounted with Vectashield with DAPI (Vector Lab, Peterborough, UK).

    Techniques: Expressing, Fluorescence, Mouse Assay, Injection, Western Blot, Binding Assay, Staining, Molecular Weight

    Alms1 − / − Retinal Structure and Function following Subretinal Delivery of Triple AAV Vectors (A) Immunohistochemical (IHC) analysis with anti-3xflag antibodies of retinal cryosections from C57BL/6J mice 2 months following subretinal injections of either triple AAV2/8 encoding for ALMS1 under the control of the PR-specific GRK1 promoter (top panels) or PBS (bottom panels). The pictures are representative of n = 4 injected eyes. The scale bar (20 μm) is depicted in the figure. Merge, overlay of DAPI and ALMS1. ONL, outer nuclear layer; OS, outer segment. (B) Spectral domain optical coherence tomograms (SD-OCT) analysis of Alms1 − / − mice injected subretinally with either triple GRK1- ALMS -AAV vectors (white bars) in one eye or PBS in the contralateral eye (black bars). Results are reported as means ± SE. (C and D) Electroretinographic analysis of a-wave (C) and b-wave (D) light responses of Alms1 − / − mice injected subretinally with either triple GRK1- ALMS -AAV vectors (white bars) or PBS in the contralateral eye (black bars). Results are reported as means ± SE. Light intensity of 20 cd s/m 2 (a-wave); background white light of 50 cd s/m 2 and light intensity of 20 cd s/m 2 (b-wave).

    Journal: Molecular Therapy

    Article Title: Triple Vectors Expand AAV Transfer Capacity in the Retina

    doi: 10.1016/j.ymthe.2017.11.019

    Figure Lengend Snippet: Alms1 − / − Retinal Structure and Function following Subretinal Delivery of Triple AAV Vectors (A) Immunohistochemical (IHC) analysis with anti-3xflag antibodies of retinal cryosections from C57BL/6J mice 2 months following subretinal injections of either triple AAV2/8 encoding for ALMS1 under the control of the PR-specific GRK1 promoter (top panels) or PBS (bottom panels). The pictures are representative of n = 4 injected eyes. The scale bar (20 μm) is depicted in the figure. Merge, overlay of DAPI and ALMS1. ONL, outer nuclear layer; OS, outer segment. (B) Spectral domain optical coherence tomograms (SD-OCT) analysis of Alms1 − / − mice injected subretinally with either triple GRK1- ALMS -AAV vectors (white bars) in one eye or PBS in the contralateral eye (black bars). Results are reported as means ± SE. (C and D) Electroretinographic analysis of a-wave (C) and b-wave (D) light responses of Alms1 − / − mice injected subretinally with either triple GRK1- ALMS -AAV vectors (white bars) or PBS in the contralateral eye (black bars). Results are reported as means ± SE. Light intensity of 20 cd s/m 2 (a-wave); background white light of 50 cd s/m 2 and light intensity of 20 cd s/m 2 (b-wave).

    Article Snippet: Seventy-two hours post-infection, cells were washed once with PBS, fixed for 7 min with 4% paraformaldehyde (PFA) in PBS, washed three times with PBS, and mounted with Vectashield with DAPI (Vector Lab, Peterborough, UK).

    Techniques: Immunohistochemistry, Mouse Assay, Injection

    Expression of Rit2 mRNA and protein in the mouse retina. The top panels show immunohistochemistry of retinal flat-mounts ( A - D ) and retinal sections ( E - H ). Images are staining with anti-RIT2 antibody ( A , E ), anti-POU4F2 antibody ( B , F ), merged image of RIT2 and POU4F2 ( C , G ), and nuclear staining by 4', 6-diamidino-2-phenylindole (DAPI; D , H ). RIT2-positive cells are more numerous than POU4F2-positive RGCs. E : RIT2 protein is detected in the ganglion cell layer (GCL), inner plexiform layer (IPL), and cells in the inner nuclear layer (INL). Short arrowheads point to RIT2-positive cells located at the inner border of the INL; long arrowheads point to RIT2-positive cells at the outer aspect of the INL. Scale bar: 50 μm. I : This panel shows Rit2 mRNA expression. Laser capture microdissection was used to collect tissues of GCL, INL, and outer nuclear layer (ONL) from mouse retinal sections, and RNA of each layer was analyzed by reverse transcription-quantitative polymerase chain reaction. The mRNA level of Rit2 , Pou4f1 , and Pou4f2 was normalized by that of Gapdh . J : RIT2 protein levels were analyzed at different ages. Protein lysates were prepared from mouse retinas at postnatal day 2 (P2), P5, P15, and adult (8 weeks old), and analyzed by western blotting with anti-RIT2 antibody and anti-α tubulin antibody for control. Protein signals were visualized by chemiluminescence with X-ray films. K : This panel shows relative quantification of RIT2 protein levels. Western blot signals on the X-ray films were scanned and analyzed with ImageJ software. The signal intensity of RIT2 was normalized by that of α tubulin and presented as the RIT2/tubulin ratio. RIT2 protein levels increased from P2 to adults by more than 15 fold.

    Journal: Molecular Vision

    Article Title: RIT2, a neuron-specific small guanosine triphosphatase, is expressed in retinal neuronal cells and its promoter is modulated by the POU4 transcription factors

    doi:

    Figure Lengend Snippet: Expression of Rit2 mRNA and protein in the mouse retina. The top panels show immunohistochemistry of retinal flat-mounts ( A - D ) and retinal sections ( E - H ). Images are staining with anti-RIT2 antibody ( A , E ), anti-POU4F2 antibody ( B , F ), merged image of RIT2 and POU4F2 ( C , G ), and nuclear staining by 4', 6-diamidino-2-phenylindole (DAPI; D , H ). RIT2-positive cells are more numerous than POU4F2-positive RGCs. E : RIT2 protein is detected in the ganglion cell layer (GCL), inner plexiform layer (IPL), and cells in the inner nuclear layer (INL). Short arrowheads point to RIT2-positive cells located at the inner border of the INL; long arrowheads point to RIT2-positive cells at the outer aspect of the INL. Scale bar: 50 μm. I : This panel shows Rit2 mRNA expression. Laser capture microdissection was used to collect tissues of GCL, INL, and outer nuclear layer (ONL) from mouse retinal sections, and RNA of each layer was analyzed by reverse transcription-quantitative polymerase chain reaction. The mRNA level of Rit2 , Pou4f1 , and Pou4f2 was normalized by that of Gapdh . J : RIT2 protein levels were analyzed at different ages. Protein lysates were prepared from mouse retinas at postnatal day 2 (P2), P5, P15, and adult (8 weeks old), and analyzed by western blotting with anti-RIT2 antibody and anti-α tubulin antibody for control. Protein signals were visualized by chemiluminescence with X-ray films. K : This panel shows relative quantification of RIT2 protein levels. Western blot signals on the X-ray films were scanned and analyzed with ImageJ software. The signal intensity of RIT2 was normalized by that of α tubulin and presented as the RIT2/tubulin ratio. RIT2 protein levels increased from P2 to adults by more than 15 fold.

    Article Snippet: The flat mounts were washed with 0.1% PBST, mounted in VECTASHIELD HardSet Mounting Medium with 4', 6-diamidino-2-phenylindole (H-1500, Vector Laboratories, Burlingame, CA), and examined on an LSM 510 inverted laser scanning confocal microscope (Carl Zeiss, Thornwood, NY).

    Techniques: Expressing, Immunohistochemistry, Staining, Laser Capture Microdissection, Real-time Polymerase Chain Reaction, Western Blot, Software