Overview

  • Product name

    Anti-Nucleophosmin antibody
    See all Nucleophosmin primary antibodies
  • Description

    Rabbit polyclonal to Nucleophosmin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    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.

  • Positive control

    • HeLa or mouse fibroblast extract.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 99% PBS, 0.1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
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).

Target

  • Function

    Involved 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 disease

    A 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 similarities

    Belongs to the nucleoplasmin family.
  • Post-translational
    modifications

    Acetylated at C-terminal lysine residues, thereby increasing affinity to histones.
    ADP-ribosylated.
    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 localization

    Nucleus, 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
  • Database links

  • Alternative names

    • B23 antibody
    • MGC104254 antibody
    • NO38 antibody
    • NPM antibody
    • NPM_HUMAN antibody
    • NPM1 antibody
    • Nucleolar phosphoprotein B23 antibody
    • Nucleolar protein NO38 antibody
    • Nucleophosmin (nucleolar phosphoprotein B23 numatrin) antibody
    • Nucleophosmin antibody
    • nucleophosmin nucleoplasmin family member 1 antibody
    • Nucleophosmin/nucleoplasmin family member 1 antibody
    • Numatrin antibody
    • OTTHUMP00000161024 antibody
    • OTTHUMP00000161025 antibody
    • OTTHUMP00000223397 antibody
    • OTTHUMP00000223398 antibody
    see all

Images

  • 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 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).
  • 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.

    See Abreview

  • 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.

References

This product has been referenced in:

  • Malfatti MC  et al. APE1 and NPM1 protect cancer cells from platinum compounds cytotoxicity and their expression pattern has a prognostic value in TNBC. J Exp Clin Cancer Res 38:309 (2019). Read more (PubMed: 31307523) »
  • Ho SM  et al. Bisphenol A and its analogues disrupt centrosome cycle and microtubule dynamics in prostate cancer. Endocr Relat Cancer 24:83-96 (2017). Read more (PubMed: 27998958) »
See all 19 Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (Saos)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Saos
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Feb 17 2016

Application
Western blot
Sample
Human Purified protein (Concentrated secreted serum free medium)
Loading amount
15 µg
Specification
Concentrated secreted serum free medium
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Aparna Mitra

Verified customer

Submitted Jul 11 2008

Application
Western blot
Sample
Mouse Cell lysate - nuclear (Mouse embryonic stem cells)
Loading amount
20 µg
Specification
Mouse embryonic stem cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 05 2007

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (embryonic stem cells)
Specification
embryonic stem cells
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10%

Abcam user community

Verified customer

Submitted Mar 01 2006

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Primary hepatocytes)
Specification
Primary hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10

Ms. Louise Mary Treanor

Verified customer

Submitted Nov 14 2005

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