Analysis Note

ControlIncluded Positive Antigen Control:
Catalog # 12-305, 3T3/A31 cell lysate. Add 2.5 µL of 2-mercapto-ethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research CategorySignaling

Immunoprecipitation:
4 µg of a previous lot of antibody immunoprecipitated paxillin from 500 µg of a 3T3/A31 RIPA lysate.

Immunocytochemistry:
5-10 µg/mL of a previous lot gave positive immunostaining for paxillin in REF (rat embryo fibroblast) 52 cells fixed with 3.7% paraformaldehyde for 5 minutes.

Anti-Paxillin Antibody, clone 5H11 is an antibody against Paxillin for use in IC, IP & WB.

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

Paxillin, a focal adhesion scaffolding protein, is phosphorylated on multiple tyrosine residues in response various stimulations including integrin stimulation, growth factor stimulation, and oncogenic transformation (v-Src, v-Crk and BCR/ABL). Paxillin binds many other focal adhesion proteins including FAK, Vinculin, Talin, and Tensin. As such, Paxillin and it tyrosine phosphorylation, help regulate the dynamics of the focal adhesions and the actin cytoskeleton which results in cell morphology and cell migration characteristics of the cell. Paxillin is most notable phosphorylated on tyrosines 31 and 118 (Tyr31 and Tyr118). Interestingly, Vinculin and FAK bind to overlapping sites on Paxillin. Additionally, there is an increase in Paxillin phosphorylation at Tyr118 in vinculin (-/-) and Vinculin Y822F cells. Paxillin Y31F Y118F expression restores apoptosis and inhibits Erk activity.

Immunogen

GST fusion protein containing fulllength human paxillin. Clone 5H11.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-581

Other Notes

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

Physical form

Protein G Purified

Purified mouse monoclonal IgG1 in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Store at -20°C.

Format: Purified

Quality

Routinely evaluated by Western Blot on RIPA lysates from 3T3/A31 fibroblasts.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected paxillin in RIPA lysates from 3T3/A31 fibroblasts.

Specificity

This antibody recognizes paxillin.

Other species cross-reactivity not tested.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon first thaw, 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. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Target description

68 kDa