Validated for:

• Western blot-WB {{RESOURCE:PL327|Customer Testimonial}}
• Immunofluorescence-IF
• Immunoprecipitation-IP
• Enzyme-linked immunosorbent assay-ELISA
• Immunohistochemistry-IHC

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 is purified by protein G affinity chromatography.

Host/isotype/clone:

• Mouse/IgG2b
• Clone: 27B10.4

Reactivity:

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

Amount of material:

• APY03: 2 x 100 µl when reconstituted (120-150 µg per tube)
• APY03-S: 1 x 25 µl when reconstituted (30-37.5 µg per tube)

Working concentrations:

• WB 1:500 dilution 2.4-3.0 µg/ml final 100 ml working volume - full size Ab
• IF 1:1000 1.2-1.5 µg/ml final 200 ml working volume - full size Ab
• IP 5 µl neat 6-7.5 µg per IP 40 assays
• ELISA 1:10,000 diln 0.12-0.15 µg/ml final 1000 ml working volume-full size Ab
• IHC 1:125 dilution 9.6-12 µg/ml final 25 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

Sensitivity:

• Detects ≤10 ng phosphotyrosine labeled BSA

Specificity:

• Highly selective: Chemically phosphotyrosinated BSA (10 ng) is easily detected in WB, while 1 µg of non-phosphotyrosinated BSA is not—demonstrating a ≥100-fold specificity.
• In native samples, it detects multiple species in lysates treated with a tyrosine phosphatase inhibitor (vanadate) and shows no reactivity to a lysate treated with a serine/threonine phosphatase inhibitor (calyculin A).
• Demonstrates comparable sensitivity and specificity to the widely used 4G10 antibody in unbiased performance comparisons {{RESOURCE:PL327|Customer Testimonial}}