Anti-Phospho-Bad (S99) Antibody

Product Description

Specifications

Size	400 µL
Estimated Lead Time	1-2 weeks
Species	Human
Host Species	Rabbit
Accession Number	Q92934
Gene Id	572
Gene Symbols	BAD
Protein Name / Synonyms	Phospho-Bad(S99)
Clonality	Polyclonal
Preparation	This Bad Antibody is generated from rabbits immunized with a KLH conjugated synthetic phosphopeptide corresponding to amino acid residues surrounding S99 of human Bad.
Concentration (lot specific)	0.500 mg/ml
Recommended Applications	IF, Dot Blotting, IHC-P
Research Area	Autophagy
Shipping Type	Blue ice
Storage	-20°C

Description

Introduction

Bad is a member of the BCL-2 family. BCL-2 family members are known to be regulators of programmed cell death. This protein positively regulates cell apoptosis by forming heterodimers with BCL-xL and BCL-2, and reversing their death repressor activity. Proapoptotic activity of this protein is regulated through its phosphorylation. Protein kinases AKT and MAP kinase, as well as protein phosphatase calcineurin are found to be involved in the regulation of this protein. Bad is phosphorylated on one or more of Ser-75, Ser-99, Ser-118 and Ser-134 in response to survival stimuli, which blocks its pro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75 promotes heterodimerization with 14-3-3 proteins. This interaction then facilitates the phosphorylation at Ser-118, a site within the BH3 motif, leading to the release of Bcl-X(L) and the promotion of cell survival. Ser-99 is the major site of AKT/PKB phosphorylation, Ser-118 the major site of protein kinase A (CAPK) phosphorylation.

Specificity

Predicted Reactivity

Mouse, Rat

Other Names

BAD; BBC6; BCL2L8; Bcl2 antagonist of cell death; Bcl-2-binding component 6; Bcl-2-like protein 8; Bcl-XL/Bcl-2-associated death promoter

NCBI Accession #

NP_004313.1;NP_116784.1

Other Accession #

O35147; Q61337

Format

Type

Peptide Affinity Purified Rabbit Polyclonal Antibody (Pab)

Calculated Molecular Weight (Da)

18392

Recommended Dilutions

IF: 1:200; DB: 1:500; IHC: 1:50-100

	Fluorescent confocal image of HeLa cells stained with phospho-Bad-S99 antibody. HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with 102-15272 phospho-Bad-S99 primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor^® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 µg/ml, 5 min). Note the highly specific localization of the phospho-Bad-S99 mainly to the cytoplasm.
	Dot blot analysis of anti-hBad-pS99 Phospho-specific Pab (Cat# 102-15272) on nitrocellulose membrane. 50ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5ug per ml.
	Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.

Antigen Source

HUMAN

Storage/Stability

2-8°C (short-term); -20°C (long-term)

Expiration:

12 months from the date of shipment when stored properly.

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