To analyze human AGXT2 transgene expression cell slides with primary aortic cells were fixed with 1:1 acetone-methanol solution for 10 min at 4 °C, washed 3 × 2 min with ice-cold PBS and blocked with Dako Protein Blocking solution for 20 min at room temperature. Afterwards, chamber slides were incubated with 1:100 rabbit polyclonal anti-FLAG antibodies (Sigma-Aldrich, St. Louis, MI, USA, Catalog #7425) for 2 h at 37 °C, washed 3 × 2 min with PBS and subsequently incubated with 1:250 anti-rabbit antibodies coupled with fluorescence dye (The Jackson Laboratory, Bar Harbor, ME, USA) at room temperature for 1 h. Finally, slides were washed 3 × 2 min with PBS, stained with 1:1000 DAPI and mounted with Moviol. To demonstrate the expression of human AGXT2-FLAG transgene in endothelial cells 1:100 rat anti-CD31 antibodies (Biolegend, San Diego, CA, USA, Catalog # 102401) (as marker of endothelial cells33 (link)), and 1:250 anti-rat secondary antibodies (The Jackson Laboratory, Bar Harbor, ME, USA) were used. Double staining of CD31 and FLAG-tagged transgene was performed in the same way.
Anti rat secondary antibodies
Manufactured by Jackson ImmunoResearch
Sourced in United States
Anti-rat secondary antibodies are reagents used to detect the presence of rat primary antibodies in various experimental techniques, such as Western blotting, immunohistochemistry, and ELISA. These antibodies are designed to specifically bind to the Fc region of rat immunoglobulins, allowing for the visualization and quantification of target proteins in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti cd31 antibodies, by BioLegend (2 mentions)
Anti rabbit antibodies, by Jackson ImmunoResearch (2 mentions)
Anti flag antibody, by Merck Group (2 mentions)
Protein blocking solution, by Agilent Technologies (2 mentions)
Fluorescence dye, by Jackson ImmunoResearch (1 mentions)
4 6 diamidino 2 phenylindole dapi, by BD (1 mentions)
Guinea pig anti insulin, by Agilent Technologies (1 mentions)
3 protocols using anti rat secondary antibodies
1
Visualization of AGXT2 Transgene Expression
2
Immunohistochemical Analysis of Pancreatic Tissues
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Tissues were fixed in 4% paraformaldehyde at 4°C for 4 hours, followed by cryoprotection in 30% sucrose overnight before snap-freezing. Slides of 6µm-thickness was performed during sectioning of the samples. Primary antibodies used in this study were: Guinea pig anti-insulin (DAKO, Carpinteria, CA, USA) and rat polyclonal anti-CD31 (Becton-Dickinson Biosciences). The secondary antibodies were Cy3-conjugated anti-guinea pig or anti-rat secondary antibodies generated from donkey (Jackson ImmunoResearch Labs, West Grove, PA, USA). Nucleus staining was performed with 4,6-Diamidino-2-phenylindole (DAPI, Becton-Dickinson Biosciences, San Jose, CA, USA).
3
Visualizing AGXT2 Transgene Expression
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To analyze human AGXT2 transgene expression cell slides with primary aortic cells were xed with 1:1 acetone-methanol solution for 10 minutes at 4°C, washed 3x2 minutes with ice-cold PBS and blocked with Dako Protein Blocking solution for 20 minutes at room temperature. Afterwards, chamber slides were incubated with 1:100 rabbit polyclonal anti-FLAG antibodies (Sigma-Aldrich, St. Louis, MI, USA, Catalog #7425) for 2 hours at 37°C, washed 3x2 minutes with PBS and subsequently incubated with 1:250 antirabbit antibodies coupled with uorescence dye (The Jackson Laboratory, Bar Harbor, ME, USA) at room temperature for 1 hour. Finally, slides were washed 3x2 minutes with PBS, stained with 1:1,000 DAPI and mounted with Moviol. To demonstrate the expression of human AGXT2-FLAG transgene in endothelial cells 1:100 rat anti-CD31 antibodies (Biolegend, San Diego, CA, USA, Catalog # 102401) (as marker of endothelial cells [33] ), and 1:250 anti-rat secondary antibodies (The Jackson Laboratory, Bar Harbor, ME, USA) were used. Double staining of CD31 and FLAG-tagged transgene was performed in the same way.
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