Analysis Note

ControlIncluded positive antigen control of non-stimulated A431 lysate (Cat. No. 12-301). Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

This Anti-Src Antibody, clone N6L is validated for use in WB for the detection of Src.

Research CategoryCell Structure

Research Sub CategoryCytoskeletal Signaling

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

pp60c-Src is the prototype non-receptor tyrosine kinase. Homology to Src in three regions is shared by a number of proteins: SH1 being the catalytic domain, SH2 being a phospho-tyrosine binding domain, and SH3 being a proline-binding domain. The Src SH2 domain binds to phospho-Tyr527 (phosphorylated by Csk) within the Src molecule, causing Src to adopt an inactive conformation. Mutation of Tyr527 to phenylalanine removes the SH2-binding target and results in a constitutively active kinase. A second mutation at the conserved lysine residue at the ATP-binding site abolishes kinase activity, but without Tyr527, the conformation of the protein is open, allowing interaction with endogenous activators. Consequently, this double-mutant exhibits a dominant-negative phenotype

Immunogen

N-terminal 6His-tagged recombinant full-length human Src expressed by baculovirus in Sf21 insect cells. Purified using Ni2+/NTA agarose.

Linkage

Replaces: 05-889

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Cultured supernantant containing 0.05% sodium azide.

Quality

Western Blot Analysis: A 1:500 to 1:1000 dilution of this antibody detected Src in RIPA lysates from A431 cells.

Specificity

Recognizes Src. Crossreacts with Yes and weakly with Fyn. No crossreactivity observed with Lck or Lyn.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~ 60 kDa