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Paxillin Mouse Monoclonal Antibody

Cat Number: MAB-80123
Conjugate: Unconjugated
Size: 100 ug
Clone: PXC-10
Concentration: Adding 500ul of PBS buffer will yield a concentration of 100 ug/500ul.
Host: Mouse
Isotype: IgG1
Immunogen: C-terminal part of recombinant chicken paxillin (amino acids 305-559).
Reactivity: Bovine, Chicken, Hamster, Human, Mouse, Rat
Applications:

Immunocytochemistry: 1ug/ml
Western blot: 1-2ug/ml

Molecular Weight: 65kDa
Purification: Ascites
Background:

The paxillin gene can be alternatively spliced to include 1 of 2 alternative exons, generating beta and gamma isoforms. Paxillin is a 68-kDa focal adhesion protein that is phosphorylated on tyrosine residues in fibroblasts in response to transformation by v-src, treatment with platelet-derived growth factor, or cross-linking of integrins. The 68-kD protein (paxillin) is a cytoskeletal component that localizes to the focal adhesions at the ends of actin stress fibers in chicken embryo fibroblasts. It is also present in the focal adhesions of Madin-Darby bovine kidney (MDBK) epithelial cells but is absent, like talin, from the cell-cell adherens junctions of these cells.

Form: Liquid
Buffer: Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as preservative.
Storage: Store at -20˚C for one year from date of receipt
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Western blot analysis of Paxillin using anti-Paxillin antibody.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The
sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A431 whole cell lysates
Lane 2: human Hela whole cell lysates
Lane 3: human MCF-7 whole cell lysates
Lane 4: human CACO-2 whole cell lysates
Lane 5: rat liver tissue lysates
Lane 6: rat RH35 whole cell lysates
Lane 7: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes.Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Paxillin antigen affinity purified monoclonal antibody at 1 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Paxillin at approximately 65 kDa. The expected band size for Paxillin is at 65 kDa.