Bmi1 Recombinant Rabbit mAb (S-3313-71)

Catalog Number: S0B6771 Application: WB, IHC-P, ICC, ChIP Reactivity: Hu Conjugation: Unconjugated Brand: Starter

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$100 USD

Size: 25μl 100μl

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Product Details

Product Specification

Host	Rabbit
Antigen	BMI-1
Immunogen	Synthetic Peptide
Location	Cytoplasm, Nucleus
Accession	P35226
Clone Number	S-3313-71
Antibody Type	Recombinant mAb
Isotype	IgG
Application	WB, IHC-P, ICC, ChIP
Reactivity	Hu
Positive Sample	A549, HeLa, U-2 OS, Molt-4
Predicted Reactivity	Bv, Ct
Purification	Protein A
Concentration	0.5 mg/ml
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
Stability & Storage	12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution

application	dilution	species
WB	1:1000	Hu
IHC-P	1:200	Hu
ICC	1:500	Hu
ChIP	1:10	Hu

Background

Bmi1 (B-cell-specific Moloney murine leukemia virus integration site 1) is a critical polycomb group protein that functions as an essential component of the Polycomb Repressive Complex 1 (PRC1), playing a pivotal role in epigenetic gene silencing by catalyzing the monoubiquitination of histone H2A at lysine 119, thereby repressing key target genes such as the INK4a/ARF locus (which encodes tumor suppressors p16INK4a and p14ARF) to regulate cell cycle progression, cellular senescence, and stem cell self-renewal; beyond its normal physiological functions in development and hematopoiesis, Bmi1 is frequently overexpressed in various malignancies, where it drives tumorigenesis, promotes cancer stem cell maintenance, confers resistance to chemotherapy and radiation, and serves as a significant prognostic marker and potential therapeutic target across numerous cancer types including breast, lung, and colorectal cancers.

Picture

Western Blot

WB result of Bmi1 Recombinant Rabbit mAb
Primary antibody: Bmi1 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: A549 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: U-2 OS whole cell lysate 20 µg
Lane 4: Molt-4 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 37 kDa
Observed MW: 40 kDa

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human tonsil. Anti-Bmi1 antibody was used at 1/200 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human colon. Anti-Bmi1 antibody was used at 1/200 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human colon cancer. Anti-Bmi1 antibody was used at 1/200 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human lung adenocarcinoma. Anti-Bmi1 antibody was used at 1/200 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Immunocytochemistry

ICC shows positive staining in HeLa cells. Anti-Bmi1 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).

ChIP

Chromatin immunoprecipitation (ChIP) was performed on NCCIT cells cross- linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication.
Parallel reactions used Bmi1 Recombinant Rabbit mAb (S-3313-71) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:10 for immunoprecipitation.
Post-immunoprecipitation, both samples were washed, eluted, and cross-links reversed. Purified DNA was analyzed by qPCR.

qPCR showed the enrichment of HOXA2 and SAT-α in BMI-1 Recombinant Rabbit
mAb (S-3313-71)-immunoprecipitated sample.