Overview

  • Product name

    Mouse Nucleophosmin ELISA Kit
    See all Nucleophosmin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Plasma 8 3.9%
    Inter-assay
    Sample n Mean SD CV%
    Plasma 3 4.8%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    600 pg/ml
  • Range

    1.56 ng/ml - 100 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 105 103% - 108%
    Cell culture extracts 95 91% - 98%
    Heparin Plasma 108 105% - 112%
    EDTA Plasma 114 113% - 116%
    Citrate Plasma 107 103% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Cow, Human
  • Product overview

    Nucleophosmin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse Nucleophosmin protein in serum, plasma, cell culture supernatant, and cell extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:


    Samples in Sample Diluent NS: 860 pg/mL


    Samples in 1X Cell Extraction Buffer PTR: 600 pg/mL

  • Notes

    Mouse Nucleophosmin is involved in an array of diverse cellular processes, including ribosome biogenesis, protein chaperoning, histone assembly, cell proliferation, and regulation of tumor suppressors p53/TP53 and ARF. In concert with BRCA2, Nucleophosmin also regulates centrosome duplication. Mouse Nucleophosmin is 292 aa in length and shares 99% and 95% sequence homology with rat and human Nucleophosmin, respectively.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Nucleophosmin Capture Antibody 1 x 600µl
    10X Mouse Nucleophosmin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Mouse Nucleophosmin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
  • Alternative names

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

Applications

Our Abpromise guarantee covers the use of ab216172 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Nucleophosmin were measured in duplicates, interpolated from the Nucleophosmin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nucleophosmin concentration was determined to be 21.07 ng/mL in neat serum, 26.17 ng/mL in neat plasma (citrate), 21.49 ng/mL in neat plasma (EDTA), and 44.43 ng/mL in neat plasma (heparin).

  • The concentrations of Nucleophosmin were measured in duplicates, interpolated from the Nucleophosmin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nucleophosmin concentration was determined to be 134.31 ng/mL in neat serum, 138.54 ng/mL in neat plasma (citrate), 138.47 ng/mL in neat plasma (EDTA), and 150.88 ng/mL in neat plasma (heparin).

  • The concentrations of Nucleophosmin were measured in duplicates, interpolated from the Nucleophosmin standard curves and corrected for sample dilution. Undiluted samples are as follows: RAW 264.7 media 2.5%, L929 media 5%, and J774A.1 media 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nucleophosmin concentration was determined to be 2,767.43 ng/mL in neat RAW 264.7 media, 1,790.80 ng/mL in neat L929 media, and 889.01 ng/mL in neat J774A.1 media. RAW 264.7 media was cultured in HGDMEM media with kanamycin and L-glutamine for 24 hours (serum free) and then for another 48 hours with 1% PHA. J774A.1 media was cultured for 72 hours in HGDMEM media with kanamycin, 10% fetal bovine serum, and 1.5% PHA plus 10 ng/mL PMA. L929 media was cultured for 72 hours in MEM media with kanamycin, 10% horse serum, and 1.5% PHA plus 10 ng/mL PMA.

  • The concentrations of Nucleophosmin were measured in duplicate and interpolated from the Nucleophosmin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nucleophosmin concentration was determined to be 57.49 ng/mL in RAW 264.7 cell extract and 100.26 ng/mL in C6 cell extract.

     

  • The concentrations of Nucleophosmin were measured in duplicates, interpolated from the Nucleophosmin standard curves and corrected for sample dilution. Undiluted samples are as follows: L929 unstimulated media 5% and L929 stimulated media 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nucleophosmin concentration was determined to be 3.12 ng/mL in neat unstimulated L929 media, and 92.60 ng/mL in neat stimulated L929 media. L929 media was cultured for 72 hours in MEM media with kanamycin and 10% horse serum without (unstimulated) and with (stimulated) 1.5% PHA plus 10 ng/mL PMA.

Protocols

References

ab216172 has not yet been referenced specifically in any publications.

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