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PhosphoPlus® Elk-1 (Ser383) Antibody Kit #9180

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
Phospho-Elk-1 (Ser383) Antibody #9181	100 µl	W	H (M) (R) (Mk) (Z) (Dg)	47	Rabbit
Elk-1 Antibody #9182	100 µl	W	H M R Z	47	Rabbit
Elk-1 Control Proteins #9183	150 µl			40 kDa nonphosphorylated, 50 kDa phosphorylated	E. Coli
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat
Anti-biotin, HRP-linked Antibody #7075	100 µl				Goat
Biotinylated Protein Ladder Detection Pack #7727	100 µl
20X LumiGLO^® Reagent and 20X Peroxide #7003	5 ml each

Applications Key: W=Western Blotting
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey Z=Zebrafish Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Elk-1 Antibody detects endogenous levels of total Elk-1 protein. Phospho-Elk-1 (Ser383) Antibody detects EIk-1 protein only when activated by phosphorylation at serine 383.

Western Blotting

Western analysis of Elk-1 fusion protein expressed from E. coli with or without phosphorylation with purified Erk2 enzyme using Phospho-Elk-1 (Ser383) #9181 (upper) or Elk-1 Antibodies #9182 (lower).

Description

The PhosphoPlus® Elk-1 (Ser383) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of Elk-1. Bacterially expressed Elk-1 serves as a negative control. The same protein phosphorylated by p42 MAP Kinase (Erk2) serves as a positive control.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide corresponding to residues surrounding Ser383 of human Elk-1, or with a synthetic peptide derived from the sequence of human Elk-1. Antibodies are purified by protein A and peptide affinity chromatography.

Background

The transcription factor Elk-1 is a component of the ternary complex that binds the serum response element (SRE) and mediates gene activity in response to serum and growth factors (1-3). Elk-1 is phosphorylated by MAP kinase pathways at a cluster of S/T motifs at its carboxy terminus; phosphorylation at these sites, particularly Ser383, is critical for transcriptional activation by Elk-1. Elk-1 appears to be a direct target of activated MAP kinase: (a) biochemical studies indicate that Elk-1 is a good substrate for MAP kinase; (b) the kinetics of Elk-1 phosphorylation and activation correlate with MAP kinase activity; (c) interfering mutants of MAP kinase block Elk-1 activation in vivo. Other studies have shown that Elk-1 (Ser383) is also a target of the stress-activated kinase SAPK/JNK (4,5).

Application References

Aplin, A. E. et al. (2001) Integrin-mediated adhesion regulates ERK nuclear translocation and phosphorylation of Elk-1. J. Biol. Chem. 153, 273-282. Applications: IC-ABC Western Blotting
Hu, Y. et al. (2000) E-selectin-dependent signaling via the mitogen-activated protein kinase pathway in vascular endothelial cells. J. Immunol. 165, 2142-2418. Applications: Western Blotting
Wang, D. and Richmond, A. (2001) Nuclear factor-kappa B activation by the CXC chemokine melanoma growth-stimulatory activity/growth-regulated protein involves the MEKK1/p38 mitogen-activated protein kinase pathway. J. Biol. Chem. 276, 3650-3659. Applications: Western Blotting

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Protocols

9181:: Western Blotting
9182:: Western Blotting

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.