HeLa cells were cultured under serum-free conditions for 24 hours and either left untreated or stimulated with 10 ng/mL EGF for various times (as indicated). Cell lysates were then prepared with RIPA buffer and Shc was immunoprecipitated with a polyclonal anti-Shc antibody. The immunoprecipitates were resolved by SDS-PAGE and transferred to PVDF. The membrane was blocked with a casein/Tween 20 buffer, then incubated with mouse anti-Shc [pS36] antibody (Catalog no. 4828M) at 1.0 µg/mL. After washing, the membrane was incubated with an anti-mouse HRP-conjugated secondary antibody and signals were detected using an ECL detection method.
|Tested species reactivity||Dog, Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Phosphopeptide containing amino acid sequence ELP[pS]PSA, conjugated to KLH.|
|Storage buffer||PBS, pH 7.3, with sucrose, PEG|
|Contains||0.09% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody specifically recognizes the Shc/p66 isoform when it is phosphorylated at serine 36.
A recommended positive control for Western blot is EGF-treated HepG2 cells.
Signaling adapter that couples activated growth factor receptors to signaling pathways. Participates in a signaling cascade initiated by activated KIT and KITLG/SCF. Isoform p46Shc and isoform p52Shc, once phosphorylated, couple activated receptor tyrosine kinases to Ras via the recruitment of the GRB2/SOS complex and are implicated in the cytoplasmic propagation of mitogenic signals. Isoform p46Shc and isoform p52Shc may thus function as initiators of the Ras signaling cascade in various non-neuronal systems. Isoform p66Shc does not mediate Ras activation, but is involved in signal transduction pathways that regulate the cellular response to oxidative stress and life span. Isoform p66Shc acts as a downstream target of the tumor suppressor p53 and is indispensable for the ability of stress-activated p53 to induce elevation of intracellular oxidants, cytochrome c release and apoptosis. The expression of isoform p66Shc has been correlated with life span . Participates in signaling downstream of the angiopoietin receptor TEK/TIE2, and plays a role in the regulation of endothelial cell migration and sprouting angiogenesis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.