Detects proteins containing phosphorylated tyrosine residues in Western blots. In Western blots of pervanadate-treated cell lysates, clone 216954 binds Phospho-Tyrosine in a broad manner largely independent of the surrounding amino acid sequence. No cross-reactivity with proteins or peptides containing phosphorylated serine or threonine residues is observed.
Monoclonal Rat IgG2A Clone # 216954
Protein A or G purified from hybridoma culture supernatant
KLH-coupled Phospho-Tyrosine synthetic peptide
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Phospho-Tyrosine by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 50 μM pervanadate (PV) for 15 minutes. PVDF membrane was probed with 1 µg/mL of Rat Anti-Phospho-Tyrosine Monoclonal Antibody (Catalog # MAB16761), followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). Tyrosine-phosphorylated proteins were detected (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 10.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Phosphorylation of tyrosine residues in signaling proteins by protein tyrosine kinases mediates a variety of cellular processes, including cell growth, differentiation, adhesion, motility, death, and metabolism. Dysregulation of tyrosine phosphorylation has been implicated in the development of many human diseases, such as diabetes and cancer. Antibodies specific for phospho-tyrosine have been invaluable reagents in the studies of signaling pathways initiated by tyrosine phosphorylation.
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