Phospho-14-3-3 ζ/δ (Ser58)/η (Ser59)/γ (Ser59)/β/α (Ser60) (E6B3G) Rabbit mAb (#52631) Datasheet Without Images Cell Signaling Technology

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

Applications:

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Rabbit IgG

UniProt ID:

#P61981, #P31946, #Q04917, #P63104

Entrez-Gene Id:

7532, 7529, 7533, 7534

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, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-14-3-3 ζ/δ (Ser58)/η (Ser59)/γ (Ser59)/β/α (Ser60) (E6B3G) Rabbit mAb detects endogenous levels of 14-3-3 ζ/δ only when phosphorylated at Ser58, 14-3-3 η and 14-3-3 γ only when phosphorylated at Ser59, and 14-3-3 α/β only when phosphorylated at Ser60. This antibody does not cross-react with 14-3-3 ε, 14-3-3 θ, or 14-3-3 σ. This antibody also recognizes a non-specific band of unknown origin at 80 kDa and 180 kDa.

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology

Monkey, Chicken, Bovine, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser58 of human 14-3-3 ζ/δ protein.

Background

The 14-3-3 family of proteins plays a key regulatory role in signal transduction, checkpoint control, apoptotic and nutrient-sensing pathways (1,2). 14-3-3 proteins are highly conserved and ubiquitously expressed. There are at least seven isoforms, β, γ, ε, σ, ζ, τ, and η that have been identified in mammals. The initially described α and δ isoforms are confirmed to be phosphorylated forms of β and ζ, respectively (3). Through their amino-terminal α helical region, 14-3-3 proteins form homo- or heterodimers that interact with a wide variety of proteins: transcription factors, metabolic enzymes, cytoskeletal proteins, kinases, phosphatases, and other signaling molecules (3,4). The interaction of 14-3-3 proteins with their targets is primarily through a phospho-Ser/Thr motif. However, binding to divergent phospho-Ser/Thr motifs, as well as phosphorylation independent interactions has been observed (4). 14-3-3 binding masks specific sequences of the target protein, and therefore, modulates target protein localization, phosphorylation state, stability, and molecular interactions (1-4). 14-3-3 proteins may also induce target protein conformational changes that modify target protein function (4,5). Distinct temporal and spatial expression patterns of 14-3-3 isoforms have been observed in development and in acute response to extracellular signals and drugs, suggesting that 14-3-3 isoforms may perform different functions despite their sequence similarities (4). Several studies suggest that 14-3-3 isoforms are differentially regulated in cancer and neurological syndromes (2,3).
Phosphorylation of 14-3-3ζ at Ser58 may regulate dimerization and affect its ability to interact with partner proteins, including p53 and ASK1 (6,7).

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

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

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