B23/NPM1 Recombinant antibody

• Featured Product
• KD/KO Validated

B23/NPM1 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, IP, ELISA

Conjugate

Unconjugated

CloneNo.

1G4

Cat no : 82030-1-RR

Synonyms

B23, NPM1, Numatrin, Nucleophosmin, NPM

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Validation Data Gallery

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  WB analysis using 82030-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 82030-1-RR (B23 antibody) at dilution of 1:40000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 82030-1-RR

  WB result of B23/NPM1 antibody (82030-1-RR; 1:100000; incubated at room temperature for 1.5 hours) with sh-Control and sh-B23/NPM1 transfected HepG2 cells.

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  IP experiment of Jurkat using 82030-1-RR

  IP result of anti-B23/NPM1 (IP:82030-1-RR, 4ug; Detection:82030-1-RR 1:8000) with Jurkat cells lysate 1560 ug.

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  IHC staining of human prostate cancer using 82030-1-RR

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 82030-1-RR (B23/NPM1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human prostate cancer using 82030-1-RR

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 82030-1-RR (B23/NPM1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human normal colon using 82030-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 82030-1-RR (B23/NPM1 antibody) at dilution of 1:8000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 82030-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using B23 antibody (82030-1-RR, Clone: 1G4 ) at dilution of 1:1000 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  Affinity and Kinetic Characterization of 82030-1-RR

  Biolayer interferometry (BLl) kinetic assays of 82030-1-RR against Human B23/NPM1 were performed. The affinity constant is 0.409 nM.

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Tested Applications

• Positive WB detected in: HEK-293 cells, HeLa cells, Jurkat cells, THP-1 cells
• Positive IP detected in: Jurkat cells
• Positive IHC detected in: human prostate cancer tissue, human normal colon; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:8000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

82030-1-RR targets B23/NPM1 in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: B23/NPM1 fusion protein Ag0286
• Full Name: nucleophosmin (nucleolar phosphoprotein B23, numatrin)
• Calculated Molecular Weight: 33 kDa
• Observed Molecular Weight: 38 kDa
• GenBank Accession Number: BC002398
• Gene Symbol: NPM1
• Gene ID (NCBI): 4869
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Nucleophosmin (NPM1,B23) is a putative ribosome assembly factor with a high affinity for peptides containing nuclear localization signals (NLSs). The transport of proteins across the nuclear envelope is a selective, multistep process involving several cytoplasmic factors. Proteins must be recognized as import substrates, dock at the nuclear pore complex and translocate across the nuclear envelope in an ATP-dependent fashion. Several cytosolic and nuclear proteins that are central to this process have been identified. The 38 kDa nuclear protein nucleophosmin is involved in ribosomal assembly and rRNA transport. It is an abundant protein that is highly phosphorylated by Cdc2 kinase during mitosis.