Anti-Paxillin (phospho Y118) antibody

• WB

Supplier Data

Western blot - Anti-Paxillin (phospho Y118) antibody (AB4833)

A 66 kDa band corresponding to Paxillin pTyr118 was enhanced across treated cell lines. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.

All lanes:

Western blot - Anti-Paxillin (phospho Y118) antibody (ab4833) at 1/500 dilution

Lane 1:

COS-7 (African green monkey kidney fibroblast-like cell line) whole cell lysate at 30 µg

Lane 2:

COS-7 whole cell lysate treated with 100 uM Pervanadate for 1 hour at 30 µg

Lane 3:

A431 (Human epidermoid carcinoma cell line) whole cell lysate at 30 µg

Lane 4:

A431 whole cell lysate treated with 100 uM Pervanadate for 1 hour at 30 µg

Lane 5:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 30 µg

Lane 6:

HeLa whole cell lysate treated with 100 uM Pervanadate for 1 hour at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/5000 dilution

Predicted band size: 65 kDa

false

• WB

Supplier Data

Western blot - Anti-Paxillin (phospho Y118) antibody (AB4833)

Peptide Competition : Extracts were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4^oC.

Prior incubation with :

1- no peptide,

2- the non-phosphopeptide corresponding to the immunogen

3- a generic phosphotyrosine containing peptide

4- the phosphopeptide immunogen

Bands were detected using the Tropix WesternStar method.

The data show that only the peptide corresponding to ab4833 blocks the antibody signal, thereby demonstrating the specificity of the antibody.

All lanes:

Western blot - Anti-Paxillin (phospho Y118) antibody (ab4833) at 0.75 µg/mL

All lanes:

NMµMG (mouse mammary) cells transfected with EGFP-tagged paxillin

Secondary

All lanes:

goat F(ab’)2 anti-rabbit IgG alkaline phosphatase

Predicted band size: 65 kDa

false