Product nameAnti-Paxillin (phospho Y31) antibody
See all Paxillin primary antibodies
DescriptionRabbit polyclonal to Paxillin (phospho Y31)
Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Chicken
Synthetic peptide (Human) derived from the region of human paxillin that contains tyrosine 31.
- ICC/IF NIH/3T3 treated with PDGF or TGF-beta. WB: NM_MG cells transfected with the wild-type EGFP-tagged paxillin, A431 cell lysate treated with pervanadate, HeLa cell lysate treated with TNF alpha, NIH/3T3 cell lysate treated with PDGF, and A549 cell lysate treated with PMA.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA
Concentration information loading...
PurityImmunogen affinity purified
Purification notesPurified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated paxillin, and ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine, irrespective of the sequence. The final product is generated by affinity chromatography using a paxillin-derived peptide that is phosphorylated at tyrosine 31.
Our Abpromise guarantee covers the use of ab4832 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration.
Please see Grace et al., 2003.
|WB||1/1000. Predicted molecular weight: 68 kDa.|
FunctionCytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion).
Sequence similaritiesBelongs to the paxillin family.
Contains 4 LIM zinc-binding domains.
modificationsPhosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens.
Cellular localizationCytoplasm > cytoskeleton. Cell junction > focal adhesion.
- Information by UniProt
- FLJ16691 antibody
- FLJ23042 antibody
- Paired box protein Pax 1 antibody
Immunocytochemistry/ Immunofluorescence of NIH/3T3 (mouse embryo fibroblast cell line) treated with 50 ng of PDGF for 10 minutes with ab4832 (1/250). A Goat anti-Rabbit IgG (H+L) Alexa Fluor® 488 conjugate (1/2000) was used as a secondary (Green). DAPI (Blue) was used as a counter stain. F-actin was stained with Alexa Fluor® 555 Rhodamine Phalloidin (1/300). Panel e is untreaded cells and panel f had no primary antibody.
All lanes : Anti-Paxillin (phospho Y31) antibody (ab4832)
Lane 1 : A431 (human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) whole cell lysate treated for 1 Hr with 100 µM of Pervanadate
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated for 20 minutes with 50 ng/mL of TNF alpha
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 6 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate treated for 10 minutes with 50 ng/mL of PDGF
Lane 7 : A549 (human lung carcinoma cell line) whole cell lysate
Lane 8 : A549 (human lung carcinoma cell line) whole cell lysate treated for 20 minutes with 200 nM of PMA
Lysates/proteins at 20 µl per lane.
All lanes : Goat anti-Rabbit IgG (H+L) HRP conjugate at 1/2500 dilution
Developed using the ECL technique.
Predicted band size: 68 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
ab4832 was diluted between 1/500 and 1/2000.
Extracts prepared from NM
µMG cells transfected with either the wild-type EGFP-tagged paxillin or a site directed mutant, in which tyrosine 31 was changed to phenylalanine, were resolved on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with 1% BSA, followed by incubation with 0.5 µg/mL anti-paxillin [pY31] antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected by chemiluminescence using the Tropix WesternStar detection method and Kodak BioMax ultraclear film. The data show detection of paxillin phosphorylation with the wild-type but not the Y31F mutant recombinant proteins. Note that the recombinant EGFP-tagged paxillin proteins run at a higher Mr as opposed to the endogenous protein and that in both cell types that the antibody recognizes the endogenous wild-type paxillin protein, which migrates at the expected Mr of 68 kDa. Ext
Colocalization of phospho Paxillin and actin in TGF-beta treated NM
µMG cells. Green = actin stress fibers, Orange = Paxillin [pY31], Yellow = colocalization.
This product has been referenced in:
- Gill MB et al. KSHV-TK is a tyrosine kinase that disrupts focal adhesions and induces Rho-mediated cell contraction. EMBO J 34:448-65 (2015). Read more (PubMed: 25471072) »
- Kawada I et al. Paxillin mutations affect focal adhesions and lead to altered mitochondrial dynamics: relevance to lung cancer. Cancer Biol Ther 14:679-91 (2013). IHC . Read more (PubMed: 23792636) »