711633

antibody from Invitrogen Antibodies

Targeting: GCGR GGR

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711633

Provider

Invitrogen Antibodies

Product name

Glucagon Receptor Recombinant Polyclonal Antibody (21HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Horse, Bovine, Dog

Antibody clone number

21HCLC

Concentration

0.5 mg/mL

Western blot

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) and HEK-293 (Lane 5). The blots were probed with Anti-Glucaogon Receptor/ GCGR Recombinant Rabbit Polyclonal Antibody (Product # 711633, 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 # NP0321BOX), 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, HepG2 cells were fixed and permeabilized for detection of endogenous Glucagon Receptor/GCGR using Anti- Glucagon Receptor/GCGR Recombinant Rabbit Polyclonal Antibody (Product # 711633, 5µg/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 localisation of Glucagon Receptor/GCGR. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.