Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Chromatin Immunoprecipitation (ChIP), CyTOF-ready

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 384515
View all BMI-1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human BMI-1
Asp96-Gly326
Accession # P35226

Specificity

Detects human BMI-1 in direct ELISAs  and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human BMI-1 Antibody (384515)

Detection of Human BMI‑1 antibody by Western Blot.

Detection of Human BMI‑1 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. Gels were loaded with 30 µg of whole cell lysate (WCL), 20 µg of cytoplasmic (Cyto), and 10 µg of nuclear extracts (Nuc). PVDF membrane was probed with 0.1 µg/mL Mouse Anti-Human BMI-1 Monoclonal Antibody (Catalog # MAB33341) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for BMI-1 was detected at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Detection of BMI-1-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of BMI‑1-regulated Genes by Chromatin Immunoprecipitation.

HeLa human cervical epithelial carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. BMI-1/DNA complexes were immunoprecipitated using 5 µg Mouse Anti-Human BMI-1 Monoclonal Antibody (Catalog # MAB33341) or control antibody (Catalog # MAB003) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Mouse IgG Secondary Antibody (Catalog # BAF007). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thehoxc13promoter was detected by standard PCR.
Detection of BMI‑1 antibody in HeLa Human Cell Line antibody by Flow Cytometry.

Detection of BMI‑1 in HeLa Human Cell Line by Flow Cytometry.

HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human BMI-1 Monoclonal Antibody (Catalog # MAB33341, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Applications for Human BMI-1 Antibody (384515)

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: HeLa human cervical epithelial carcinoma cell line chromatin, hoxc13 promoter detected by standard PCR

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HeLa human cervical epithelial carcinoma cell line fixed with paraformaldehyde and permeabilized with saponin

Western Blot

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Reviewed Applications

Read 1 review rated 5 using MAB33341 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: BMI-1

BMI-1 (B cell-specific Moloney-MLV integration site #1) is a 45 kDa protooncogene that is a class II member of the Polycomb group of genes. It participates in the formation of a large multimeric complex termed PRC1 that inhibits target gene transcription. Loss of BMI-1 function precludes stem cells from self-replicating. Human BMI-1 contains an N-terminal RING-finger domain (aa 17-56), an NLS (aa 81-95) and a C-terminal Pro/Ser-rich region (aa 251-326). Human BMI-1 shares 99%, 97%, 99% and 99% aa sequence identity with bovine, mouse, feline and canine BMI-1, respectively.

Long Name

B Lymphoma Mo-MLV Insertion Region 1

Alternate Names

BMI1, PCGF4, RNF51

Entrez Gene IDs

648 (Human); 12151 (Mouse)

Gene Symbol

BMI1

UniProt

P35226

Additional BMI-1 Products

Product Documents for Human BMI-1 Antibody (384515)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human BMI-1 Antibody (384515)

For research use only

Citations for Human BMI-1 Antibody (384515)

Customer Reviews for Human BMI-1 Antibody (384515) (1)

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  • Human BMI-1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MCF-7 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 03/22/2017
    MCF-7 cell lysate probed with 1:2,500 Mouse anti-Human BMI-1 Antibody. Buffer: 5% BSA in PBS. Secondary Ab: anti-Mouse IgG 1:5,000.
    Human BMI-1 Antibody (384515) MAB33341

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Protocols

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