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Human, reacts against Thr199-phosphorylated NPM/B23
Immunogen:
Modified peptide
Formulation:
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 50% glycerol. Bovine serum albumin is present at 1 mg/ml final concentration (USA origin).
Preparation:
The antibody was purified by antigen-affinity chromatography.
Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 10 µl per 5 ml antibody dilution buffer for each mini-gel. For IHC, use a 1:50 dilution of antibody for staining. Antigen retrieval using 0.01 M sodium citrate buffer is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Hela cells were treated with 300 μM mimosine for 16 h, then placed in complete media (lane 1) or media containing 200 ng/ml nocodazole (lane 2) for an additional 18 h. Nuclear extract was western blotted with Poly6192. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.
Formalin-fixed paraffin-embedded human thymus tissue was stained with Poly6192 and developed with an alkaline phosphatase chromogen substate (red color). Tissue was counterstained with H&E (blue/pink). Magnification, 40X.
NPM/B23 (also known as numatrin, nucleophosmin 1, nucleolar phosphoprotein B23) is a member of the nucleophosmin/nucleoplasmin family that shuttles between the nucleus and cytoplasm. This protein regulates the stability and transcriptional activity of p53 and acts as a molecular chaperone. NPM/B23 preferentially binds to denatured proteins and has been shown to stimulate DNA polymerase activity and control the duplication of centrosomes. The chaperone activity of NPM/B23 is regulated by protein kinase CK2 and promotes release of denatured substrates from NPM/B23. NPM/B23 is modified by phosphorylation onThr199. This protein has been shown to bind to RNA and DNA and interact with nucleolar proteins including nucleolin, protein P120, HIV-1 Rev protein, and hepatitis delta antigens. The Poly6192 antibody recognizes human phosphorylated NPM/B23 (Thr199) and has been shown to be useful for Western blotting.
Nucleolar protein, can shuttle between nucleus and cytoplasm
Function:
Regulates the stability and transcriptional activity of p53, acts as a molecular chaperone. Preferentially binds to denatured proteins. Has been shown to stimulate DNA polymerase activity and control the duplication of centrosomes
Regulation:
Chaperone activity is regulated by protein kinase CK2, promotes release of denatured substrates from NPM/B23
Modification:
Phosphorylation
Interaction:
Binds to RNA, DNA, and interacts with nucleolar proteins including nucleolin, protein P120, HIV-1 Rev protein, and hepatitis delta antigens
Antigen References:
1. Szebeni A, et al. 2003. J. Biol. Chem. 278:9107. 2. Colombo E, et al. 2002. Nat. Cell Biol. 4:529. 3. Chan PK, et al. 1986. J. Biol. Chem. 261:14335.
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Purified anti-NPM/B23-Phosphorylated (Thr199)
Hela cells were treated with 300 μM mimosine for 16 h, then placed in complete media (lane 1) or media containing 200 ng/ml nocodazole (lane 2) for an additional 18 h. Nuclear extract was western blotted with Poly6192. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.
Formalin-fixed paraffin-embedded human thymus tissue was stained with Poly6192 and developed with an alkaline phosphatase chromogen substate (red color). Tissue was counterstained with H&E (blue/pink). Magnification, 40X.
