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Synthetic peptide corresponding to the C-terminal 15 residues of rat Glut4 cross-linked to Keyhole limpet hemocyanin (Peptide available as ab115831.)
Our Abpromise guarantee covers the use of ab654 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000 - 1/2500. Detects a band of approximately 45 kDa (predicted molecular weight: 54.8 kDa).Can be blocked with Rat Glucose Transporter GLUT4 peptide (ab115831).
The samples MUST NOT be boiled before running the gel as highly hydrophobic membrane proteins, like glucose transporters, aggregate severely at high temperatures due to the hydrophobic effect, which increases as a direct function of temperature. If the samples are boiled, much of the transporter will aggregate so severely it doesn't enter the running gel and higher order oligomers will be observed.
It is recomended to use 3% milk block.
|IP||Use at an assay dependent concentration. PubMed: 18234908|
|Immunomicroscopy||Use at an assay dependent concentration. Can also can be used for Immunogold EM (purified IgG). See references for specific protocols.|
|IHC-P||Use at an assay dependent concentration.|
ICC/IF image of ab654 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 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 (ab654, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
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