Products:Signal Transduction >> Protein Phosphorylation >> Tyrosine Kinases >> Receptor Tyrosine Kinases
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ab5644 has been referenced in 6 publications.
Publishing research using ab5644? Please let us know so that we can cite the reference in this datasheet
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Peptide Competition: Extracts prepared from NIH3T3 cells expressing EGFR were starved for 30 hours, then stimulated for 10 minutes with 30 ng/mL EGF (+), or left unstimulated (-), then resolved by SDS-PAGE on a 6% Tris-glycine gel, and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 µg/mL ab5644 antibody, following prior incubation with: no peptide (1, 2), the phosphopeptide immunogen (3, 4), or, the non phosphopeptide corresponding to the phosphopeptide immunogen (5, 6). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, demonstrating the specificity of the ab5644 antibody for this phosphorylated residue. The data also show the activation of the EGFR after stimulation with EGF.
All lanes : Anti-EGFR (phospho Y1068) antibody (ab5644) at 1/200 dilution
Lane 1 : A431 whole cell lysate - not treated
Lane 2 : A431 whole cell lysate - 100ng/ml EGF for 1 minute
Lane 3 : A431 whole cell lysate - 100ng/ml EGF for 2.5 minutes
Lane 4 : A431 whole cell lysate - 100ng/ml EGF for 5 minutes
Lane 5 : A431 whole cell lysate - 100ng/ml EGF for 10 minutes
Lane 6 : A431 whole cell lysate - 100ng/ml EGF for 20 minutes
Lane 7 : A431 whole cell lysate - 100ng/ml EGF for 40 minutes
Secondary
HRP conjugated Goat anti-rabbit antibody
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 2 minutes
This image is courtesy of an Abreview submitted by Mr Samir Nuseibeh
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