RabMAb

Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

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Key features and details

  • Rabbit monoclonal [EP1848Y] to Nucleophosmin - BSA and Azide free
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free
    See all Nucleophosmin primary antibodies

  • Description

    Rabbit monoclonal [EP1848Y] to Nucleophosmin - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Nucleophosmin aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, MEF, PC-12, 3T3-L1 cell lysates; IHC-P: Human glioma, breast carcinoma and cervical carcinoma tissues; ICC/IF: HeLa cells; Flow Cyt: HeLa cells.

  • General notes

    ab239847 is the carrier-free version of ab52644 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab239847 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EP1848Y

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab239847 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
WB Use at an assay dependent concentration. Detects a band of approximately 35,38 kDa (predicted molecular weight: 33 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.

    • 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

      see all

    • 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 - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Western blot - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Anti-Nucleophosmin antibody [EP1848Y] (ab52644) at 1/20000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 33 kDa
      Observed band size: 250,27,33 kDa
      why is the actual band size different from the predicted?



      The band at 250kDa might be an oligomer and the band at 33kDa might be an isoform of NPM.

      Blocking/Diluting buffer: 5% NFDM/TBST

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    • Flow Cytometry - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Flow Cytometry - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

      Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Nucleophosmin with Purified ab52644 at 1:40 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling Nucleophosmin with purified ab52644 at 1/300 dilution (1.14 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    • Immunocytochemistry/ Immunofluorescence - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Immunocytochemistry/ Immunofluorescence - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Nucleophosmin with purified ab52644 at 1/100 dilution (5 µg/mL). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

      ab52644, unpurified, showing positive staining in human breast carcinoma tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

      ab52644, unpurified, showing positive staining in human cervical carcinoma tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] - BSA and Azide free (ab239847)

      ab52644, unpurified, showing positive staining in human glioma tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52644).

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab239847? Please let us know so that we can cite the reference in this datasheet.

    ab239847 has not yet been referenced specifically in any publications.

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