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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- 702602 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Glucagon Receptor Recombinant Rabbit Monoclonal Antibody (21H1L9)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Horse, Bovine, Dog
- Antibody clone number
- 21H1L9
- Concentration
- 0.5 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of U-87 MG (Lane 1), Hep G2 (Lane 2), COLO 205 (Lane 3), HeLa (Lane 4), HEK-293 (Lane 5) and PC-12 (Lane 6). The blots were probed with Anti-Glucaogon Receptor/ GCGR Recombinant Rabbit Monoclonal Antibody (Product # 702602, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 50 kDa band corresponding to Glucaogon Receptor/ GCGR was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of U-87 MG (Lane 1), Hep G2 (Lane 2), COLO 205 (Lane 3), HeLa (Lane 4), HEK-293 (Lane 5) and PC-12 (Lane 6). The blots were probed with Anti-Glucaogon Receptor/ GCGR Recombinant Rabbit Monoclonal Antibody (Product # 702602, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 50 kDa band corresponding to Glucaogon Receptor/ GCGR was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, U87-MG cells were fixed and permeabilized for detection of endogenous Glucagon Receptor/GCGR using Anti- Glucagon Receptor/GCGR Recombinant Rabbit Monoclonal Antibody (Product # 702602, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Glucagon Receptor/GCGR protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating Membrane localization of Glucagon Receptor/GCGR. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
