Product nameAnti-Nucleophosmin antibody
See all Nucleophosmin primary antibodies
DescriptionRabbit polyclonal to Nucleophosmin
Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Rat, Human
This product was produced with the following immunogens:
Synthetic peptide corresponding to Human Nucleophosmin aa 23-38.
Synthetic peptide corresponding to Human Nucleophosmin aa 226-240.
- HeLa or mouse fibroblast extract.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.1% BSA
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab15440 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||1/100. Used at a dilution of 1/100 (see Abreview for further information).|
|WB||1/500 - 1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 33 kDa).|
FunctionInvolved in diverse cellular processes such as ribosome biogenesis, centrosome duplication, protein chaperoning, histone assembly, cell proliferation, and regulation of tumor suppressors p53/TP53 and ARF. Binds ribosome presumably to drive ribosome nuclear export. Associated with nucleolar ribonucleoprotein structures and bind single-stranded nucleic acids. Acts as a chaperonin for the core histones H3, H2B and H4. Stimulates APEX1 endonuclease activity on apurinic/apyrimidinic (AP) double-stranded DNA but inhibits APEX1 endonuclease activity on AP single-stranded RNA. 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, regulates centrosome duplication. Regulates centriole duplication: phosphorylation by PLK2 is able to trigger centriole replication. Negatively regulates the activation of EIF2AK2/PKR and suppresses apoptosis through inhibition of EIF2AK2/PKR autophosphorylation. Antagonizes the inhibitory effect of ATF5 on cell proliferation and relieves ATF5-induced G2/M blockade (PubMed:22528486).
Involvement in diseaseA chromosomal aberration involving NPM1 is found in a form of non-Hodgkin lymphoma. Translocation t(2;5)(p23;q35) with ALK. The resulting chimeric NPM1-ALK protein homodimerize and the kinase becomes constitutively activated.
A chromosomal aberration involving NPM1 is found in a form of acute promyelocytic leukemia. Translocation t(5;17)(q32;q11) with RARA.
A chromosomal aberration involving NPM1 is a cause of myelodysplastic syndrome (MDS). Translocation t(3;5)(q25.1;q34) with MLF1.
Defects in NPM1 are associated with acute myelogenous leukemia (AML). Mutations in exon 12 affecting the C-terminus of the protein are associated with an aberrant cytoplasmic location.
Sequence similaritiesBelongs to the nucleoplasmin family.
modificationsAcetylated at C-terminal lysine residues, thereby increasing affinity to histones.
Phosphorylated at Ser-4 by PLK1 and PLK2. Phosphorylation at Ser-4 by PLK2 in S phase is required for centriole duplication and is sufficient to trigger centriole replication. Phosphorylation at Ser-4 by PLK1 takes place during mitosis. Phosphorylated by CDK2 at Ser-125 and Thr-199. Phosphorylation at Thr-199 may trigger initiation of centrosome duplication. Phosphorylated by CDK1 at Thr-199, Thr-219, Thr-234 and Thr-237 during cell mitosis. When these four sites are phosphorated, RNA-binding activity seem to be abolished. May be phosphorylated at Ser-70 by NEK2. The Thr-199 phosphorylated form has higher affinity for ROCK2. CDK6 triggers Thr-199 phosphorylation when complexed to Kaposi's sarcoma herpesvirus (KSHV) V-cyclin, leading to viral reactivation by reducing viral LANA levels.
Sumoylated by ARF.
Cellular localizationNucleus, nucleolus. Nucleus, nucleoplasm. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Generally nucleolar, but is translocated to the nucleoplasm in case of serum starvation or treatment with anticancer drugs. Has been found in the cytoplasm in patients with primary acute myelogenous leukemia (AML), but not with secondary AML. Can shuttle between cytoplasm and nucleus. Co- localizes with the methylated form of RPS10 in the granular component (GC) region of the nucleolus. Colocalized with nucleolin and APEX1 in nucleoli. Isoform 1 of NEK2 is required for its localization to the centrosome during mitosis.
- Information by UniProt
- B23 antibody
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ab15440 detecting Nucleophosmin in interphase mouse primary embryonic fibroblasts. Cells were also counterstained with anti gamma tubulin antibody and DAPI in order to highlight the centrossomes and the nucleus (respectively). Secondary antibodies: goat anti-rabbit IgG conjugated to Alexa 488 (for ab15440) and goat anti-mouse IgG conjugated to Alexa 594 (for gamma-tubulin antibody).
ab15440 detecting Nucleophosmin in mitotic mouse primary embryonic fibroblasts. Cells were also counterstained with anti gamma tubulin antibody and DAPI in order to highlight the centrossomes and the nucleus (respectively). Secondary antibodies: goat anti-rabbit IgG conjugated to Alexa 488 (for ab15440) and goat anti-mouse IgG conjugated to Alexa 594 (for gamma-tubulin antibody).
Western blot analysis of various cell lines (25ug/lane) labeling Neucleophosmin with ab15440 at 1/500. A HRP-conjugated goat anti-rabbit IgG (1/20000) was used as the secondary antibody.
ab15440 staining human purified nucleophosmin from concentrated secreted serum free medium. 15 µg of purified protein were loaded in to each lane. The primary antibody was diluted 1/500 and incubated with the blot for 15 hours at 4°C. A HRP conjugated donkey anti-rabbit IgG antibody, diluted 1/5000, was used as the secondary.
ab15440 (4µg/ml) staining nucleophosmin in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human kidney tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/500 with a rabbit polyclonal antibody recognizing Nucleophosmin (ab15440) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
This product has been referenced in:
- Holley CL et al. Cytosolic accumulation of small nucleolar RNAs (snoRNAs) is dynamically regulated by NADPH oxidase. J Biol Chem 290:11741-8 (2015). WB ; Rat . Read more (PubMed: 25792744) »
- Mayor R et al. Genome distribution of replication-independent histone H1 variants shows H1.0 associated with nucleolar domains and H1X associated with RNA polymerase II-enriched regions. J Biol Chem 290:7474-91 (2015). WB ; Human . Read more (PubMed: 25645921) »