Product nameAnti-PAG antibody
See all PAG primary antibodies
DescriptionRabbit polyclonal to PAG
Tested applicationsSuitable for: IP, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Human
Recombinant fragment, corresponding to amino acids 97-432 of Human PAG.
- Human Ramos cell line, Raji cell line Mouse spleen cells
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Concentration information loading...
Purification notesPurified from rabbit antisera by precipitation methods. Purity >95% by SDS-PAGE.
Our Abpromise guarantee covers the use of ab14989 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 51.8 kDa. Use a lysis buffer made with Tris HCl (pH 8.1) and 1%laurylmaltoside (with common protease inhibitors), mild detergents such as NP-40 do not work. Non-denaturing conditions should be used|
FunctionNegatively regulates TCR (T-cell antigen receptor)-mediated signaling in T-cells and FCER1 (high affinity immunoglobulin epsilon receptor)-mediated signaling in mast cells. Promotes CSK activation and recruitment to lipid rafts, which results in LCK inhibition. Inhibits immunological synapse formation by preventing dynamic arrangement of lipid raft proteins. May be involved in cell adhesion signaling.
Tissue specificityUbiquitously expressed. Present in germinal center B-cells, plasma cells, T-cells, monocytes and platelets (at protein level).
Phosphorylated by FYN on Tyr-317 in resting T-cells; which promotes interaction with CSK. Dephosphorylated by PTPRC/CD45 upon TCR activation; which leads to CSK dissociation. May also be dephosphorylated by PTPN11. Hyperphosphorylated in mast cells upon FCER1 activation.
Cellular localizationCell membrane. Present in lipid rafts.
- Information by UniProt
- CBP antibody
- Csk binding protein antibody
- Csk-binding protein antibody
All lanes : Anti-PAG antibody (ab14989) at 1 µg/ml
Lane 1 : Human liver tissue lysate - total protein (ab29889)
Lane 2 : Lung (Human) Tissue Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 51.8 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Additional bands at: 60 kDa. We are unsure as to the identity of these extra bands.
PAG migrates on SDS PAGE gels anomalously as an 80 kDa molecule.
1. Frozen Raji cells lysate, immunoprecipitated with ab4206
2. Fresh Raji cells lysate, immunoprecipitated with ab4206
3. Frozen Raji cells lysate, immunoprecipitated with ab14989
Reprobe with ab9527.
The immunoprecipitation used covalently bound antibody to Sepharose beads as immunosorbent. This method of precipitation obviously differs from proteinA/G-sepharose or GoatxMs -Sepharose mediated immunoprecipitation protocols.
IHC image of ab14989 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14989, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
This product has been referenced in:
- Burn GL et al. Superresolution imaging of the cytoplasmic phosphatase PTPN22 links integrin-mediated T cell adhesion with autoimmunity. Sci Signal 9:ra99 (2016). Read more (PubMed: 27703032) »
- Cai H et al. Differential transformation capacity of Src family kinases during the initiation of prostate cancer. Proc Natl Acad Sci U S A 108:6579-84 (2011). IHC-P ; Mouse . Read more (PubMed: 21464326) »