Product Name:
NPM-pT199
Product Number:
ab-pn976
Target Full Name: Nucleophosmin
Target Alias: B23; Nucleolar phosphoprotein B23; Nucleolar protein NO38; Nucleophosmin (Nucleolar phosphoprotein B23, numatrin); Nucleophosmin/B23.2; Numatrin
Product Type Specific: NPM1 phosphosite-specific antibody
Antibody Code: PN976
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T199
Protein UniProt: P06748
Protein SigNET: NPM
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human NPM
Antibody Immunogen Sequence: CSIRD(pT)PAK
Antibody Immunogen Description: Corresponds to amino acid residues S1957 to K202. T199 phosphorylation inhibits functional activity, and binding to MDM2 and RNA, and transcription. This is a major in vivo phosphorylation site in NPM1 (≥54 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). NPM1 is known to be phosphorylated at this site in vitro by CDC7 (CDC7L1), CDK1 (CDC2), CDK2, CDK6, and COT (MAP3K8).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.95 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Dog | Rat | Mouse | Platypus.
Related Product 1: NPM S4 phosphorylation antibody (Cat. No.: AB-PN975)
Scientific Background: NPM1 (Nucleophosmin; B23) is an abundant, multifunctional chaperone protein primarily located in the nucleolus, where it regulates diverse cellular processes such as ribosome biogenesis, centrosome duplication, protein chaperoning, histone assembly, cell proliferation, and regulation of tumour suppressors p53/TP53 and ARF. It interacts with and regulates ARF and p53, maintaining cellular stability. It binds ribosomes to drive ribosome nuclear export. It is associated with nucleolar ribonucleoprotein structures and binds single-stranded nucleic acids. It acts as a chaperonin for the core histones H3, H2B and H4. It stimulates APEX1 endonuclease activity on apurinic/apyrimidinic (AP) double-stranded DNA but inhibits APEX1 endonuclease activity on AP single-stranded RNA. It may exert a control of APEX1 endonuclease activity within nucleoli devoted to repair AP on rDNA and the removal of oxidized rRNA molecules. In concert with BRCA2, it regulates centrosome duplication. It controls centriole duplication, since its phosphorylation by PLK2 is able to trigger centriole replication. It negatively regulates the activation of EIF2AK2/PKR and suppresses apoptosis through inhibition of EIF2AK2/PKR autophosphorylation. It antagonizes the inhibitory effect of ATF5 on cell proliferation and relieves ATF5-induced G2/M blockade. In complex with MYC, it enhances the transcription of MYC target genes. It prevents aggregation of nucleolar proteins, and plays a key role in DNA repair and centrosome duplication. It constantly shuttles between the nucleus and cytoplasm for these functions. Mutations in NPM1 is common in 30–35% of adult Acute Myeloid Leukemia (AML) cases, and can cause the protein to delocalize to the cytoplasm. Dysfunction from mutation of NPM1, often C-terminal insertions, cause cytoplasmic localization, serving as a is a driver of leukemia. It can be overexpressed in various solid tumours, and translocations (e.g., NPM1-ALK) are involved in rare lymphomas. This description may include information annotated by UniProt and/or Google AI.