Revision 1

#5727Store 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 Mk

SENSITIVITY:

Endogenous

MW (kDa):

78

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q99759

Entrez-Gene Id:

4215

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

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

MEKK3 (D36G5) Rabbit mAb recognizes endogenous levels of total MEKK3 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro310 of human MEKK3 protein.

Background

MAP kinase kinase kinase (MEKK3 or MAP3K3) is a serine/threonine protein kinase that activates SAPK and ERK via phosphorylation and activation of their respective MAP kinase kinases, SEK and MEK1/2 (1,2). MEKK3 also stimulates MEK5 via activation of ERK5/BMK1, which is at least partly regulated by a direct interaction between MEK5 and MEKK3 via p67phox-Bem1p (PB1) protein-protein interaction domains found in both proteins (3,4). MEKK3 modulates NF-κB activation in response to a variety of agonists including TNFα, LPS, IL-1 and LPA (5-9). Despite reports showing that phosphorylation of MEKK3 at Ser526 within the activation loop is necessary for kinase activation (10-12), at least one study suggests that dual phosphorylation at Thr516 and Ser520 is required for LPA-stimulated IKKβ/NF-κB activation (13). Phosphorylation at Thr294 appears to negatively regulate MEKK3 by promoting 14-3-3β binding and inhibition of the kinase activity (12). Phosphorylation of MEKK3 at Thr294 is diminished upon treatment of cells with LPS or TNFα, further suggesting an inhibitory role for this site (12).

  1. Blank, J.L. et al. (1996) J Biol Chem 271, 5361-8.
  2. Ellinger-Ziegelbauer, H. et al. (1997) J Biol Chem 272, 2668-74.
  3. Nakamura, K. and Johnson, G.L. (2003) J Biol Chem 278, 36989-92.
  4. Nakamura, K. et al. (2010) J Biol Chem 285, 2077-89.
  5. Yang, J. et al. (2001) Nat Immunol 2, 620-4.
  6. Zhao, Q. and Lee, F.S. (1999) J Biol Chem 274, 8355-8.
  7. Samanta, A.K. et al. (2004) J Biol Chem 279, 7576-83.
  8. Huang, Q. et al. (2004) Nat Immunol 5, 98-103.
  9. Sun, W. et al. (2009) Cell Signal 21, 1488-94.
  10. Zhang, D. et al. (2006) EMBO J 25, 97-107.
  11. Fritz, A. et al. (2006) J Biol Chem 281, 6236-45.
  12. Matitau, A.E. and Scheid, M.P. (2008) J Biol Chem 283, 13261-8.
  13. Sun, W. et al. (2010) J Biol Chem 285, 7911-18.

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 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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