RecombinantRabMAb

Recombinant Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)

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Western blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
  • Western blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
  • Immunoprecipitation - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
  • Dot Blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
  • Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20172] to Nucleophosmin (citrulline R196)
  • Suitable for: WB, Dot blot, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Nucleophosmin (citrulline R196) antibody [EPR20172]
    See all Nucleophosmin primary antibodies

  • Description

    Rabbit monoclonal [EPR20172] to Nucleophosmin (citrulline R196)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Dot blot, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Mouse Nucleophosmin aa 150-250 (citrulline R196). The exact sequence is proprietary.
    Database link: Q61937

  • Positive control

    • WB: NIH/3T3 transfected with GFP-tagged PADI4 (WT) expression vector, treated with calcium chloride and Ionomycin whole cell lysate; HEK-293T transfected with GFP-tagged PADI2 (WT) expression vector, treated with calcium chloride and Ionomycin whole cell lysate; C6 transfected with GFP-tagged PADI4 (WT) expression vector, treated with calcium chloride and Ionomycin whole cell lysate. IP: HEK 293T transfected with GFP-tagged PADI4 expression vector then treated with CaCl2 and ionomycin whole cell lysate.

  • General notes

    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.

    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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR20172

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab208015 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 1/1000. Predicted molecular weight: 32 kDa.
Dot blot 1/1000.
IP 1/30.

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 (citrulline R196) antibody [EPR20172] (ab208015)
    Western blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
    All lanes : Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015) at 1/1000 dilution

    Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) transfected with a control vector containing GFP tag, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 2 : NIH/3T3 transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 32 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Western blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
    Western blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
    All lanes : Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a control vector containing GFP tag, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 2 : HEK-293T transfected with GFP-tagged PADI2 (WT) expression vector, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 3 : C6 (rat glial tumor cell line) transfected with a control vector containing GFP tag, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 4 : C6 transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 32 kDa
    Observed band size: 37 kDa why is the actual band size different from the predicted?



     

    Exposure time:  Lane 1 and 2: 5 seconds; Lane 3 and 4: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
    Immunoprecipitation - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)

    Nucleophosmin (citrulline R196) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-tagged PADI4 expression vector for 24h then treated with 10 mM CaCl2 and 10 µM ionomycin for 2h, whole cell lysate with ab208015 at 1/30 dilution. Western blot was perfromed from the immunoprecipitate using ab208015 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T transfected with GFP-tagged PADI4 expression vector for 24h then treated with 10 mM CaCl2 and 10 µM ionomycin for 2h, whole cell lysate 10μg (Input).

    Lane 2: ab208015 IP in HEK-293T transfected with GFP-tagged PADI4 expression vector for 24h then treated with 10 mM CaCl2 and 10 µM ionomycin for 2h, whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208015 in HEK-293T transfected with GFP-tagged PADI4 expression vector for 24h then treated with 10mM CaCl2 and 10 µM ionomycin for 2h, whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST

    Exposure time : lesss than 1 second.

     

  • Dot Blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
    Dot Blot - Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)

    Dot blot analysis of Nucleophosmin (citruline R196) labeled with ab208015 at 1/1000 dilution.

    Lane 1: Nucleophosmin (citrulline R196) peptide.

    Lane 2: Nucleophosmin non-citrulline peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

  • Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)
    Anti-Nucleophosmin (citrulline R196) antibody [EPR20172] (ab208015)

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 ab208015? Please let us know so that we can cite the reference in this datasheet.

ab208015 has not yet been referenced specifically in any publications.

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