Anti-Glucose Transporter GLUT4 antibody (ab33780)

ab33780 gave a positive signal against the partial recombinant GLUT4 protein which has an expected molecular weight of 30 kDa.

IHC image of Glucose Transporter GLUT4 staining in Human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab33780, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

ab33780 staining Glucose Transporter GLUT4 in Mouse kidny tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with Acetone and blocked with 2% Horse serum for 60 minutes at 25°C. Samples were incubated with primary antibody (1/500) for 16 hours at 4°C. An Alexa Fluor®568-conjugated Donkey anti-rabbit IgG polyclonal (1/2000) was used as the secondary antibody.

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Immunohistochemical of PFA-fixed paraffin-embedded rat skeletal muscle skeletal tissue, labelling glucose transporter GLUT4 with ab33780 at a dilution of 1/200 incubated for 13 hours at 4°C in 1% BSA in TBS. Antigen retrival was with Tris-EDTA at pH 9.0 (heat mediated). Blocking was with 3% BSA incubated for 1 hour at 37°C. Secondary was a Goat anti-rabbit polyclonal Akaline Phosphotase conjugate at 1/200.

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IHC image of Glucose Transporter GLUT4 staining in Mouse normal skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab33780, 5µg/ml, for 15 mins at room temperature. A Goat anti-Rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

ab33780 staining glucose transporter GLUT4 in human heart tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/2000 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.

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