Revision 1

#13168Store 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, IF-IC

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

190, 110

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q8TCU6

Entrez-Gene Id:

57580

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:400

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.

Specificity / Sensitivity

PREX1 (D8O8D) Rabbit mAb recognizes endogenous levels of total PREX1 protein. This antibody will recognize both isoform 1 (190 kDa) and isoform 2 (110 kDa) human PREX1, but has not been observed to cross-react with human PREX2 protein.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding His770 of human PREX1 protein.

Background

Phosphoinositide-3,4,5-triphosphate (PtdIns(3,4,5)P3)-dependent Rac exchanger 1 (PREX1) is a Rac-specific GTP-exchange factor (GEF) regulated by heterotrimeric G-protein β/γ subunits and the lipid second messenger PtdIns(3,4,5)P3 (1-4). PREX1 contains two DEP (Dishevelled, Egl-10, and Pleckstrin homology) domains that coordinate heterotrimeric G-protein signaling. It also contains a Dbl-homology domain, which exhibits Rac-GEF activity, and PH and PDZ domains for interacting with upstream and downstream signaling components (1). Originally shown to modulate cellular migration of neutrophils by Rac2 activation (5-8), it is clear that PREX1 plays a broader role in modulating cell migration. PREX1 promotes metastasis of prostate cancer and melanoma cells, affects endothelial junction integrity, and is required for platelet generation and function (9-14). Research studies suggest that PREX1 plays an essential role in mediating ErbB-dependent signaling events in breast cancer by coordinating Rac activation in response to paracrine signals within the tumor microenvironment. Activation of PREX1 downstream of ErbB3 and EGFR chemokine receptors (CXCR4) promotes Rac activation, increased migration, proliferation, tumorigenesis, and metastasis in breast cancer cells (15,16). Consistent with this observation, deletion of PREX1 expression in mice results in resistance to melanoma metastasis (11). Expression of PREX1 in human tumors transplanted into mice inversely correlates with increased tumor progression and poor survival (15). Additional research studies suggest that PREX Rac-GEF activity is enhanced by phosphorylation in response to growth factors or hormones, and may require coincident dephosphorylation of two PH domain serine residues. The upstream kinases and precise regulatory mechanism remains elusive (15,17).

  1. Welch, H.C. et al. (2002) Cell 108, 809-21.
  2. Hill, K. et al. (2005) J Biol Chem 280, 4166-73.
  3. Mayeenuddin, L.H. and Garrison, J.C. (2006) J Biol Chem 281, 1921-8.
  4. Barber, M.A. et al. (2007) J Biol Chem 282, 29967-76.
  5. Welch, H.C. et al. (2005) Curr Biol 15, 1867-73.
  6. Dong, X. et al. (2005) Curr Biol 15, 1874-9.
  7. Zhao, T. et al. (2007) J Leukoc Biol 81, 1127-36.
  8. Nie, B. et al. (2010) Cell Signal 22, 770-82.
  9. Qin, J. et al. (2009) Oncogene 28, 1853-63.
  10. Wong, C.Y. et al. (2011) J Biol Chem 286, 25813-22.
  11. Lindsay, C.R. et al. (2011) Nat Commun 2, 555.
  12. Qian, F. et al. (2012) Arterioscler Thromb Vasc Biol 32, 768-77.
  13. Naikawadi, R.P. et al. (2012) Circ Res 111, 1517-27.
  14. Campbell, A.D. et al. (2013) PLoS One 8, e53982.
  15. Montero, J.C. et al. (2011) Oncogene 30, 1059-71.
  16. Sosa, M.S. et al. (2010) Mol Cell 40, 877-92.
  17. Montero, J.C. et al. (2013) Cell Signal 25, 2281-9.

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 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.
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.

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Revision 1
#13168

PREX1 (D8O8D) Rabbit mAb

Western Blotting Image 1: PREX1 (D8O8D) Rabbit mAb Expand Image
使用 PREX1 (D8O8D) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对不同细胞系的提取物进行蛋白质印迹分析。PREX1 在乳腺癌细胞中的差异表达与 Sosa, M.S. 等人描述的一致。(2010).
Immunofluorescence Image 1: PREX1 (D8O8D) Rabbit mAb Expand Image
使用 PREX1 (D8O8D) Rabbit mAb(绿色)对已经 heregulin(10 ng/ml,5 分钟;左图)或 heregulin(10 ng/ml,5 分钟)和 Wortmannin #9951(1 μM,1 小时;中间)处理的 MCF7 细胞与已经 heregulin(10 ng/ml,5 分钟;右图)处理的 MDA-MB-231 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。