Phospho-(Ser) 14-3-3 Binding Motif Antibody (#9601) Datasheet with Images Cell Signaling Technology

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

Applications:

WB, IP, IHC-P, E-P

REACTIVITY:

All

SENSITIVITY:

Endogenous

MW (kDa):

SOURCE:

Rabbit

Product Information

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunoprecipitation	1:50
Immunohistochemistry (Paraffin)	1:100
Peptide ELISA (DELFIA)	1:500

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

Phospho-(Ser) 14-3-3 Binding Motif Antibody binds peptides and proteins containing a motif composed of phospho-Ser with proline at the +2 position and arginine or lysine at the -3 position. Antibody binding is phospho-specific and largely independent of other surrounding amino acids. 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-Thr/Ser/Tyr-containing motifs. By ELISA this antibody recognizes a wide range of peptides containing the 14-3-3 binding motif, and by 2D gel Western blot analysis it recognizes a large number of presumptive 14-3-3 binding proteins.

Species Reactivity:

All Species Expected

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phospho-(Ser) 14-3-3-binding-motif containing peptides . Antibodies are purified by protein A and peptide affinity chromatography.

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 IHC-P: Immunohistochemistry (Paraffin) 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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