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Products:Signal Transduction >> Protein Phosphorylation >> Tyrosine Kinases >> Other
Overview
Product name
Anti-PAG antibody
See all PAG products (5) ...
Description
Rabbit polyclonal to PAG
Tested applications
IP, IHC-P, WBmore details
Cross reactivity
Reacts with
Mouse, Human
Immunogen
Recombinant fragment, corresponding to amino acids 97-432 of Human PAG.
Positive control
Human Ramos cell line, Raji cell line Mouse spleen cells
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 15mM Sodium Azide
Constituents: PBS, pH 7.4
Concentration
Concentration information loading...
Purity
>95% by SDS-PAGE
Purification notes
Purified from rabbit antisera by precipitation methods.
Clonality
Polyclonal
Isotype
IgG
Research areas
Signal Transduction >> Adapters >> Transmembrane
Signal Transduction >> Protein Phosphorylation >> Tyrosine Kinases >> Other
Applications
Show applications key
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 dilution.
IHC-P: Use a concentration of 5 µg/mlPerform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB: Use at an assay dependent dilution. 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)
Target
Function
Negatively 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 specificity
Ubiquitously expressed. Present in germinal center B-cells, plasma cells, T-cells, monocytes and platelets (at protein level).
Post-translational
modifications
Palmitoylated.
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 localization
Cell membrane. Present in lipid rafts.
Target information above from: UniProt accessionQ9NWQ8
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- CBP antibody
- Csk Binding Protein antibody
- Csk-binding protein antibody
see all
Database links
- Entrez Gene: 55824 Human
- Entrez Gene: 94212 Mouse
- Omim: 605767 Human
- SwissProt: Q9NWQ8 Human
- SwissProt: Q3U1F9 Mouse
- Unigene: 266175 Human
- Unigene: 23897 Mouse
Anti-PAG antibody images:
Immunoprecipitation
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.
Western blot - PAG antibody (ab14989)
All lanes : Anti-PAG antibody (ab14989) at 1 µg/ml
Lane 1 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)
Lane 2 : Lung (Human) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PAG antibody (ab14989)
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.
References for Anti-PAG antibody (ab14989)
This product has been referenced in:
- Cai Het 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) »
- Boyd RSet al. Protein profiling of plasma membranes defines aberrant signaling pathways in mantle cell lymphoma. Mol Cell Proteomics : (2009).Read more (PubMed: 19346216) »
See all 2 publications for this product
Publishing research using ab14989? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"