Revision 3
#69363
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, IF-IC
Reactivity:
H M R Mk
Sensitivity:
Endogenous
MW (kDa):
68
Source/Isotype:
Rabbit IgG
UniProt ID:
#P49023
Entrez-Gene Id:
5829
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Immunofluorescence (Immunocytochemistry) | 1:200 - 1:800 |
Storage
Specificity/Sensitivity
Phospho-Paxillin (Tyr118) (E9U9F) Rabbit mAb recognizes endogenous levels of paxillin protein only when phosphorylated at Tyr118.
Species predicted to react based on 100% sequence homology
Mouse, Rat, Chicken
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr118 of human paxillin protein.
Background
Paxillin is a multidomain protein that localizes primarily to focal adhesion sites in the extracellular matrix (1). Paxillin is one of the key components of integrin signaling, and tyrosine phosphorylation of paxillin is required for integrin-mediated cytoskeletal reorganization (2). Paxillin is phosphorylated by another focal adhesion component, focal adhesion kinase (FAK), at Tyr118 (3,4). Phospho-paxillin (Tyr118) may provide a docking site for recruitment of other signaling molecules to focal adhesions. It has been shown that the SH2 domain of Crk binds to the phosphorylated Tyr118 of paxillin (5).
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 3
Western blot analysis of extracts from HeLa cells in suspension, untreated (-) or induced attachment to fibronectin-coated plate surface (10 μg/ml, 1 hr; +), using Phospho-Paxillin (Tyr118) (E9U9F) Rabbit mAb (upper) and Paxillin (D9G12) Rabbit mAb #12065 (lower).
Immunoprecipitation of Phospho-Paxillin (Tyr118) protein from HeLa cells induced attachment to fibronectin-coated plate surface (10 μg/ml, 1 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Paxillin (Tyr118) (E9U9F) Rabbit mAb. Western blot analysis was performed using Phospho-Paxillin (Tyr118) (E9U9F) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
HeLa cells were detached with 0.04% trypsin and 0.03% EDTA, suspended in serum-free medium for one hour, and either processed in suspension (right) or plated on fibronectin-coated slides and allowed to adhere for an additional hour prior to fixation (left). Adherent cells were post-processed with λ-phosphatase to confirm phospho-specificity (center). Confocal immunofluorescent analysis was performed after staining with Phospho-Paxillin (Tyr118) (E9U9F) Rabbit mAb (green), DyLight™ 554 Phalloidin #13054 (red), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).