Revision 4

#50195Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

62, 68

Source/Isotype:

Rabbit IgG

UniProt ID:

#P49023

Entrez-Gene Id:

5829

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:800 - 1:1600

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #82351.

Specificity / Sensitivity

Paxillin (E6R6Z) Rabbit mAb recognizes endogenous levels of total paxillin protein. This antibody also recognizes α, β, and γ paxillin isoforms.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu63 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).

  1. Turner, C.E. (2000) J Cell Sci 113 Pt 23, 4139-40.
  2. Burridge, K. et al. (1992) J Cell Biol 119, 893-903.
  3. Bellis, S.L. et al. (1995) J Biol Chem 270, 17437-41.
  4. Bellis, S.L. et al. (1997) Biochem J 325 (Pt 2), 375-81.
  5. Schaller, M.D. and Parsons, J.T. (1995) Mol Cell Biol 15, 2635-45.

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

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) 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 All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

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 4
#50195

Paxillin (E6R6Z) Rabbit mAb

Western Blotting Image 1: Paxillin (E6R6Z) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Paxillin (E6R6Z) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Negative expression of paxillin protein in HL-60 cells is consistent with the predicted expression pattern.
Immunoprecipitation Image 1: Paxillin (E6R6Z) Rabbit mAb Expand Image
Immunoprecipitation of paxillin protein from PC-3 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Paxillin (E6R6Z) Rabbit mAb. Western blot analysis was performed using Paxillin (E6R6Z) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunofluorescence Image 1: Paxillin (E6R6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse lung labeled with Paxillin (E6R6Z) Rabbit mAb (green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (Alexa Fluor® 488 Conjugate) #68206 (gray pseudocolor), S6 Ribosomal Protein (54D2) Mouse mAb (Alexa Fluor® 647 Conjugate) #5548 (red), and DAPI #4083 (blue).
Immunofluorescence Image 2: Paxillin (E6R6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse pancreas labeled with Paxillin (E6R6Z) Rabbit mAb (green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (Alexa Fluor® 488 Conjugate) #68206 (gray pseudocolor), S6 Ribosomal Protein (54D2) Mouse mAb (Alexa Fluor® 647 Conjugate) #5548 (red), and DAPI #4083 (blue).
Immunofluorescence Image 1: Paxillin (E6R6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of PC-3 cells (left, high-expressing), HeLa cells (middle, low-expressing), and HL-60 cells (right, negative) using Paxillin (E6R6Z) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Immunofluorescence Image 2: Paxillin (E6R6Z) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of RAW 264.7 cells (left, positive) or Neuro-2a cells (right, negative) using Paxillin (E6R6Z) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).