Revision 1

#4515Store 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

REACTIVITY:

H M R Hm Mk

SENSITIVITY:

Endogenous

MW (kDa):

81

SOURCE:

Rabbit

UniProt ID:

#Q14155

Entrez-Gene Id:

8874

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

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

Cool1/βPix Antibody detects endogenous levels of total cool1/βPix protein. The antibody does not cross-react with cool2/αPix.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues in the center of human cool1/βPix. Antibodies are purified by peptide affinity chromatography.

Background

Cool/Pix proteins comprise a family of guanine nucleotide exchange factors (GEFs) localized to focal adhesions. The family consists of two isoforms, cool2/αpix and cool1/βPix, the latter having two splice variants that vary in their carboxy termini (1). Cool1/βPix, like other GEFs, has a DH (Dbl homology) domain, which allows binding of small GTPases and GDP/GTP exchange, and a PH (Pleckstrin homology) domain, which is important in regulating subcellular localization. Cool1/βPix also has an SH3 domain, which binds to the PAK kinase, a downstream effector of cdc42 and Rac (3,4). Phosphorylation of cool1/βPix by PAK2 downstream of MAPK signaling alters the localization of a complex containing PAK2 and cool-1/βPix, regulating formation of growth cones in response to growth factors (4). Growth factor induced activation of Rac1 via cool1/βPix was later shown to occur independently of subcellular localization (5). Endothelin-1 stimulation of mesangial cells stimulates the protein kinase A (PKA) pathway, resulting in translocation of cool-1/βPix and activation of cdc42 (6).

  1. Koh, C.G. et al. (2001) J. Cell Sci. 114, 4239-4251.
  2. Manser, E. et al. (1998) Mol. Cell 1, 183-192.
  3. Bagrodia, S. and Cerione, R.A. (1999) Trends Cell Biol. 9, 350-355.
  4. Shin, E.Y. et al. (2002) J. Biol. Chem. 277, 44417-44430.
  5. Shin, E.Y. et al. (2004) J. Biol. Chem. 279, 1994-2004.
  6. Chahdi, A. et al. (2005) J. Biol. Chem. 280, 578-584.

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

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.

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