Revision 1

#14580Store 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

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

153

SOURCE:

Rabbit

UniProt ID:

#Q99570

Entrez-Gene Id:

30849

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

PIK3R4 Antibody recognizes endogenous levels of total PIK3R4 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly825 of human PIK3R4 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

The serine/threonine PI3 kinase regulatory subunit 4 (PIK3R4, Vps15) is the mammalian homologue of the yeast vacuolar protein sorting 15 (1). PIK3R4 regulates the kinase activity of PI3K class III and anchors the kinase to cellular membranes through myristoylation (2,3). Recruitment of PI3K class III to the site of early endosome fusion and docking is directly mediated by PIK3R4 binding to the small GTPase Rab5 through its HEAT and WD-40 domains (4,5). The PIK3R4/PI3K class III plays a role in late endosome function through PIK3R4 binding to the Rab7 GTPase (6). In addition to its role in trafficking, the PIK3R4/PI3K class III complex interacts with beclin-1 to play a role during several stages of autophagy. Autophagosome formation is stimulated when Atg14 complexes with PIK3R4, PI3K class III, and beclin-1. The UVRAG protein competes with Atg14 for beclin-1 binding, forming a mutually exclusive complex with PIK3R4, PI3K class III, and beclin-1 that regulates autophagosome maturation. Autophagosome maturation is impaired in the presence of the beclin-1-binding protein Rubicon (7,8). Co-expression of PIK3R4 is required for PI3K class III activation and regulation by both beclin-1/UVRAG and by nutrients (9). Overexpression of PIK3R4 protein has been associated with decreased survival in patients with ovarian tumors, while mutations of the corresponding PIK3R4 gene are associated with metastatic melanoma, suggesting that PIK3R4 functions in cancer (10,11).

  1. Herman, P.K. et al. (1991) Cell 64, 425-37.
  2. Stack, J.H. et al. (1993) EMBO J 12, 2195-204.
  3. Panaretou, C. et al. (1997) J Biol Chem 272, 2477-85.
  4. Christoforidis, S. et al. (1999) Nat Cell Biol 1, 249-52.
  5. Murray, J.T. et al. (2002) Traffic 3, 416-27.
  6. Stein, M.P. et al. (2003) Traffic 4, 754-71.
  7. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
  8. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
  9. Yan, Y. et al. (2009) Biochem J 417, 747-55.
  10. Shull, A.Y. et al. (2012) PLoS One 7, e43369.
  11. Huang, J. et al. (2011) Genes Chromosomes Cancer 50, 606-18.

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 nonfat dry milk, 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.

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