Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
Specificity
The antibody PAG-C1 recognizes an epitope located in the intracellular C-terminal domain of Csk-binding protein (Cbp / PAG), a 46 kDa ubiquitously expressed transmembrane adaptor protein present in membrane rafts (glycosphingolipid-enriched microdomains), which however migrates on SDS PAGE gels anomalously as an 80 kDa molecule.
Cross-Reactivity (Details)
Mouse, Human, Rat, Bovine, Other not tested
Purification
Purified by protein-A affinity chromatography.
Purity
> 95 % (by SDS-PAGE)
Immunogen
C-terminal peptide (last 15 amino acids) of human Csk binding protein coupled to KLH.
Immunohistochemistry (paraffin sections): Positive tissue: appendix (germinal center of lymphatic follicle), heat-mediated antigen retrieval in citrate buffer pH 6.1. Immunoprecipitation: Positive control: RAJI human Burkitt lymphoma cell line.
Restrictions
For Research Use only
Concentration
1 mg/mL
Buffer
Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Do not freeze.
Storage
4 °C
Storage Comment
Store at 2-8°C. Do not freeze.
Davidson, Bakinowski, Thomas, Horejsi, Veillette: "Phosphorylation-dependent regulation of T-cell activation by PAG/Cbp, a lipid raft-associated transmembrane adaptor." in: Molecular and cellular biology, Vol. 23, Issue 6, pp. 2017-28, (2003) (PubMed).
Vang, Abrahamsen, Myklebust, Horejsí, Taskén: "Combined spatial and enzymatic regulation of Csk by cAMP and protein kinase a inhibits T cell receptor signaling." in: The Journal of biological chemistry, Vol. 278, Issue 20, pp. 17597-600, (2003) (PubMed).
Target
PAG1
(phosphoprotein Associated with Glycosphingolipid Microdomains 1 (PAG1))
Phosphoprotein membrane anchor with glycosphingoli,PAG (phosphoprotein associated with GEMs), also known as Cbp (Csk-binding protein), is a ubiquitously expressed 46 kDa transmembrane adaptor protein present in membrane rafts (glycosphingolipid-enriched microdomains), which however migrates on SDS PAGE gels anomalously as an 80 kDa molecule. Following tyrosine phosphorylation by Src family kinases, PAG binds and thereby activates the protein tyrosine kinase Csk, the major negative regulator of the Src family kinases. Signaling via the B-cell receptor in B cells or high affinity IgE receptor (FcepsilonRI) in mast cells leads to PAG increased tyrosine phosphorylation and Csk binding, while T cell receptor signaling causes PAG dephosphorylation, loss of Csk binding and increased activation of the protein tyrosine kinase Lck.,CBP, PAG