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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA3-048 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GPR62 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- IHC(P) analysis shows positive staining of GPR62 in human pancreatic islets.
- Concentration
- Conc. Not Determined
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GPR62 was performed by loading equal amounts of wheat germ lectin agarose bead enriched GPR receptor fractions from mock-transfected or GPR62 transfected HEK293 lysates onto a 7.5% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane, blocked and probed with a GPR62 polyclonal antibody (Product # PA3-048) at a dilution of 1:5000, overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody. Denatured GPR62 was detected at ~72kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluoresent analysis of GPR62 in HepG2 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes and blocked with 3% BSA in PBS for 30 minutes at room temperature. Cells were stained with a GPR62 rabbit polyclonal antibody (Product # PA3-048) at a dilution of 1:500 in blocking buffer for 1 hour at room temperature, and then incubated with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A-11034) at a dilution of 1:1000 for at least 30 minutes at room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of GPR62 was performed on human pancreas tissue. To expose target proteins, antigen retrieval was performed by microwaving tissues for 20 minutes in 10mM sodium citrate buffer (pH 6.0). Tissue slides were probed with a GPR62 polyclonal antibody (Product # PA3-048) at a dilution of 1:3000, overnight at 4C in a humidified chamber. Tissues were washed, and detection was performed using an ABC kit composed of biotinylated goat anti-rabbit IgG, peroxidase-conjugated avidin, and 3-amino-9-ethylcarbazole (AEC) substrate in acetate buffer. Tissues were counterstained with hematoxylin and dehydrated to prep for mounting.
