NPM1 Antibody (PD00-91)

Western Blot: NPM1 Antibody (PD00-91) [NBP3-32669] - Western blot analysis of NPM1 on different lysates with Rabbit anti-NPM1 antibody (NBP3-32669) at 1/1,000 dilution. Lane 1: Hela cell lysate Lane 2: HepG2 cell lysate Lane 3: A431 cell lysate Lane 4: C2C12 cell lysate Lane 4: RAW264.7 cell lysate Lysates/proteins at 30 ug/Lane. Predicted band size: 33 kDa Observed band size: 38 kDa Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32669) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: NPM1 Antibody (PD00-91) [NBP3-32669] - Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-NPM1 antibody (NBP3-32669) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32669) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: NPM1 Antibody (PD00-91) [NBP3-32669] - Immunocytochemistry analysis of LOVO cells labeling NPM1 with Rabbit anti-NPM1 antibody (NBP3-32669) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NPM1 antibody (NBP3-32669) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Flow Cytometry: NPM1 Antibody (PD00-91) [NBP3-32669] - Flow cytometric analysis of Daudi cells labeling NPM1. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32669, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC
• Clone: PD00-91
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

NPM1 Antibody (PD00-91) Summary

• Description: Novus Biologicals Rabbit NPM1 Antibody (PD00-91) (NBP3-32669) is a recombinant monoclonal antibody validated for use in IHC, WB, Flow and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic peptide within Human NPM1 aa 1-100 (N terminal). (Uniprot: P06748)
• Localization: Nucleus, nucleolus, nucleoplasm, cytoplasm, cytoskeleton, microtubule organizing center, centrosome.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: NPM1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:500-1:1000
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:200-1:500
  • Western Blot 1:1000
• Theoretical MW: 33 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for NPM1 Antibody (PD00-91)

• B23
• MGC104254
• NPM1
• NPMnucleophosmin/nucleoplasmin family, member 1
• Nucleolar phosphoprotein B23
• Nucleolar protein NO38
• nucleophosmin (nucleolar phosphoprotein B23, numatrin)
• Nucleophosmin
• Numatrin

Background

Nucleophosmin (NPM) is a multi-functional protein that is involved in many cellular activities including cell growth, proliferation and transformation. It is frequently overexpressed in solid tumors, and has recently been found to be mutated and aberrantly localized in the cytoplasm of leukaemic blasts in patients with acute myeloid leukemia (AML).

NMP antibodies are useful tools as tumor markers in cancer research.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: NPM1