Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb (#9606) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

WB, IP, E-P

REACTIVITY:

All

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Mouse IgG1

Product Information

Product Usage Information

Application	Dilution
Western Blotting	1:4000
Immunoprecipitation	1:20
Peptide ELISA (DELFIA)	1:1000

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

Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb binds peptides and proteins containing phospho-Ser surrounded by Pro at the +2 position and Arg/Lys at the -3 position. By ELISA, the antibody recognizes a wide range of peptides containing this phosphorylated 14-3-3 binding motif in a manner that is phospho-specific and largely independent of the surrounding amino acid sequence. The antibody weakly cross-reacts with sequences containing phospho-Thr instead of phospho-Ser in this motif, and with sequences containing phospho-Ser surrounded by Phe at the +1 position and Arg/Lys at the -3 position. No cross-reactivity is observed with corresponding nonphosphorylated sequences or with other phospho-Ser/Thr/Tyr containing motifs. Phospho-(Ser) 14-3-3 Binding Motif (4E2) Mouse mAb complements our polyclonal Phospho-(Ser) 14-3-3 Binding Motif Antibody #9601 by showing slightly different and overlapping specificity.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with phospho-(Ser) 14-3-3 binding motif peptides .

Background

The 14-3-3 proteins are a highly conserved family of proteins involved in the regulation of cell survival, apoptosis, proliferation and checkpoint control (1-5). Biological regulation by 14-3-3 is mediated through phosphorylation-dependent protein-protein interactions (6). Two different phospho-Ser-containing motifs are found within nearly all known 14-3-3 binding proteins (7). Motif 1 (Arg/Lys and Ser at positions -3 and -2, phospho-Ser at position 0, and Pro at position +2) is found in critical regulatory proteins including Bad, cdc25C, FKHRL1, PKC and c-Raf (5,7). Phospho-(Ser) 14-3-3 Binding Motif Polyclonal and (4E2) Monoclonal Antibodies provide powerful tools for the discovery and characterization of potential 14-3-3 binding proteins containing this motif and for high throughput drug discovery.

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 E-P: Peptide ELISA (DELFIA)

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

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