• Validated for Western blot-WB

Anti-phosphotyrosine antibody (APY03) was raised against a proprietary mixture of phosphotyrosine peptides conjugated to KLH. It detects a wide range of proteins that are post-translationally modified by phosphorylation of tyrosine residues. APY03 has been conjugated through its amine groups to the carbohydrate groups of horseradish peroxidase (HRP) to make AAC03-HRP. The antibody is purified via Protein G chromatography.

Host/isotype/clone:

• Mouse/IgG2b
• Clone: 27B10.4

Reactivity:

• Reactivity: All species (phosphotyrosine is identical in all species)

Amount of material:

• APY03-HRP: 1 x 100 µl when reconstituted (80-100 µg per tube)
• APY03-S: 1 x 25 µl when reconstituted (20-25 µg per tube)

Working concentrations:

• WB 1:6000 dilution 0.13-0.17 µg/ml final 600 ml working volume - full size Ab

Formulation

• Supplied as a lyophilized powder
• Buffer composition after reconstitution in 50% glycerol in water: 200 mM PIPES buffer pH7.0, 1% sucrose, 1% dextran, 50% glycerol, 10 mg/ml BSA

Sensitivity:

• Detects ≤1 ng phosphotyrosine labeled BSA

Specificity:

• Highly selective: Chemically phosphotyrosinated BSA (≤1 ng) is easily detected in WB, while 1 µg of non-phosphotyrosinated BSA is not—demonstrating a ≥1000-fold specificity.
• In native samples, it detects multiple species in lysates treated with a tyrosine phosphatase inhibitor (vanadate). It shows no reactivity to a lysate treated with a serine/threonine phosphatase inhibitor (calyculin A).