Recombinant
RabMAb

Recombinant Anti-Nucleophosmin (phospho T199) antibody [EP1857Y] (ab81551)

Overview

  • Product name

    Anti-Nucleophosmin (phospho T199) antibody [EP1857Y]
    See all Nucleophosmin primary antibodies
  • Description

    Rabbit monoclonal [EP1857Y] to Nucleophosmin (phospho T199)
  • Host species

    Rabbit
  • Specificity

    Recognizes Nucleophosmin only when phosphorylated at T199.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic phosphopeptide corresponding to residues surrounding Theonine 199 of Human Nucleophosmin

  • Positive control

    • WB: lysate from HeLa cell treated with CA; IHC-P: Human lymphoma; ICC/IF: HeLa cells.
  • General notes

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

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP1857Y
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab81551 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 1/2500 - 1/10000. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/100 - 1/250.
ICC/IF 1/100 - 1/250.
IP 1/70.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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

  • All lanes : Anti-Nucleophosmin (phospho T199) antibody [EP1857Y] (ab81551) at 1/2500 dilution

    Lane 1 : lysate from untreated HeLa cells
    Lane 2 : lysate from HeLa cells treated with CA

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled Goat anti-Rabbit at 1/2000 dilution

    Predicted band size: 33 kDa
    Observed band size: 33 kDa

  • Immunohistochemistry analysis of paraffin-embedded Human lymphoma tissue, using 1/100 ab81551.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunofluorescent staing of HeLa cells using 1/100 ab81551.
  • ab81551 staining Nucleophosmin (phospho T199) in human HeLa cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with formaldehyde, permeabilized with 0.1% Triton x100 in PBS and blocking with 5% serum was performed for 30 minutes at 230C. Samples were incubated with primary antibody (1/150 in PBS) for 1 hour at 23°C. An Alexa Fluor®488-conjugated goat polyclonal to rabbit IgG was used at dilution at 1/100 as secondary antibody.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab76539 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76539, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

References

This product has been referenced in:

  • Mukhopadhyay A  et al. 14-3-3? Prevents Centrosome Amplification and Neoplastic Progression. Sci Rep 6:26580 (2016). WB . Read more (PubMed: 27253419) »
  • Booth DG  et al. Ki-67 is a PP1-interacting protein that organises the mitotic chromosome periphery. Elife 3:e01641 (2014). ICC/IF ; Human . Read more (PubMed: 24867636) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer



Ich habe nun mehr Informationen vom Labor und kopiere diese direkt hier in die E-Mail für Sie:

"The Hela cell line was cultured in the size of 150cm²cell culture flask (#430825) from corning company. The cells were uniformly distributed in the 150cm²cell culture flask. The Hela cells were serum-starved for overnight and then the Hela cells were treated with Calyculin A 100ng/ml (work concentration) for 30min at 37C.

It’s about 5ul Calyculin A (from the stock solution at 100ug/ml) added into the 5ml cell supernatant/ medium in the flask. The cells were harvested after treatment."

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C

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Submitted Aug 12 2009

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