Revision 3
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

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

42-50

SOURCE:

Rabbit

UniProt ID:

#P61073

Entrez-Gene Id:

7852

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

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

Specificity / Sensitivity

Phospho-CXCR4 (Ser324/Ser325) Antibody recognizes endogenous levels of CXCR4 protein only when phosphorylated at Ser324 and Ser325. This antibody does not cross-react with CXCR4 protein when phosphorylated at Ser339.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Hamster, Chicken

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser324/Ser325 of human CXCR4 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

CXCR4 is a chemokine receptor that belongs to the G protein-coupled receptor family. It is activated by a small cytokine, CXCL12, also known as stromal cell derived factor 1 (SDF1) (1). The main function of CXCR4 is the mediation of the homing of progenitor cells in the bone marrow and their recruitment to sites of injury (2). More recently, CXCR4 has been studied as a potential therapeutic target in the context of autoimmune diseases (3), as well as cancer, as the receptor is involved in the regulation of migration, proliferation, and survival of cancer cells (4).

CXCR4 is rapidly phosphorylated at Ser324 and Ser325 by protein kinase C and G protein-coupled receptor kinase 6 upon treatment of cells with CXCL12 (2). Phosphorylation of CXCR4 at Ser324/Ser325 is critical to the ubiquitination and degradation of CXCR4 (6,7).

  1. Blanchet, X. et al. (2012) Front Immunol 3, 175.
  2. Döring, Y. et al. (2014) Front Physiol 5, 212.
  3. Debnath, B. et al. (2013) Theranostics 3, 47-75.
  4. Teicher, B.A. and Fricker, S.P. (2010) Clin Cancer Res 16, 2927-31.
  5. Busillo, J.M. et al. (2010) J Biol Chem 285, 7805-17.
  6. Marchese, A. and Benovic, J.L. (2001) J Biol Chem 276, 45509-12.
  7. Bhandari, D. et al. (2009) Mol Biol Cell 20, 1324-39.

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

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.
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 3
#85578

Phospho-CXCR4 (Ser324/Ser325) Antibody

Western Blotting Image 1: Phospho-CXCR4 (Ser324/Ser325) Antibody Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with constructs expressing Myc/DDK-tagged full-length human CXCR4 wild-type protein (CXCR4-WT-Myc/DDK), Myc/DDK-tagged full-length human CXCR4 S324A protein (CXCR4-S324A-Myc/DDK), Myc/DDK-tagged full-length human CXCR4 S325A protein (CXCR4-S325A-Myc/DDK), Myc/DDK-tagged full-length human CXCR4 S324A/S325A protein (CXCR4-S324A/S325A-Myc/DDK), or Myc/DDK-tagged full-length human CXCR4 S339A protein (CXCR4-S339A-Myc/DDK), using Phospho-CXCR4 (Ser324/Ser325) Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: Phospho-CXCR4 (Ser324/Ser325) Antibody Expand Image
Western blot analysis of extracts from MOLT-4 cells, untreated (-) or treated (+) with combinations of the folowing treatments as indicated: CXCL12 (100 nM, 10 min), λ-phosphatase, using Phospho-CXCR4 (Ser324/Ser325) Antibody (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower).