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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive).
Our Abpromise guarantee covers the use of ab15309 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration. PubMed: 25269858
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. See Abreview.|
|WB||Use a concentration of 0.5 µg/ml. Predicted molecular weight: 55 kDa.
The band may look broad like that for most membrane glycoproteins. A reviewer of another antibody against Glut1, ab652, suggests "do not boil sample before loading as this causes smearing of GLUT-1 bands".
ab15309 staining Glucose Transporter GLUT1 (green) in Human red blood cells tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 30 minutes at room temperature; antigen retrieval was by heat mediation in a citrate buffer, pH 6.0. Samples were incubated with primary antibody (1/500 in PBS-T + 1% PBS) for 12 hours. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Red - autofluorescence of erythrocytes.
ab15309 at a 1/100 dilution staining rat cells (neural stem cells from adult subventricular zone) by Immunocytochemistry/Immunofluorescence. The cells were incubated with the antibody for 18 hours and then bound antibody was detected using a Cy3 conjugated Goat anti-rabbit IgG (H + L).
This image is courtesy of an Abreview submitted by Martin Maurer.
ICC/IF image of ab15309 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab15309, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.