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CENPB Mouse Monoclonal Antibody [A5F6]

Usd: 350

Specification

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Catalog# HA600071

CENPB Mouse Monoclonal Antibody [A5F6]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

CENPB Mouse Monoclonal Antibody [A5F6]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human CENPB aa 1-200 / 599.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Target Molecular Weight

Predicted band size: 65 kDa

Positive Control

A431 cell lysate, Daudi cell lysate, 3T3-L1 cell lysate, PC-12 cell lysate, SH-SY5Y, human kidney tissue, human skin tissue.

Conjugation

unconjugated

Clone Number

A5F6

RRID

AB_3071688

Product Features

Form

Liquid

Concentration

2 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG2a

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:100

  • IHC-P

  • 1:100-1:500

Target

Function

This gene product is a highly conserved protein that facilitates centromere formation. It is a DNA-binding protein that is derived from transposases of the pogo DNA transposon family. It contains a helix-loop-helix DNA binding motif at the N-terminus, and a dimerization domain at the C-terminus. The DNA binding domain recognizes and binds a 17-bp sequence (CENP-B box) in the centromeric alpha satellite DNA. This protein is proposed to play an important role in the assembly of specific centromere structures in interphase nuclei and on mitotic chromosomes. It is also considered a major centromere autoantigen recognized by sera from patients with anti-centromere antibodies.

Background References

1. Otake K. et. al. CENP-B creates alternative epigenetic chromatin states permissive for CENP-A or heterochromatin assembly. J Cell Sci. 2020 Aug

2. Ohzeki JI. et. al. Human artificial chromosome: Chromatin assembly mechanisms and CENP-B. Exp Cell Res. 2020 Apr

Subcellular Location

Nucleus, centromere.

UNIPROT

SwissProt: P07199 Human

SwissProt: P27790 Mouse

Entrez Gene: 362217 Rat

Synonyms

CENP B antibody

CENP-B antibody

CENPB antibody

CENPB_HUMAN antibody

centromere autoantigen B antibody

Centromere protein B antibody

Centromere protein B, 80kDa antibody

Major centromere autoantigen B antibody

Images

  • Western blot analysis of CENPB on different lysates with Mouse anti-CENPB antibody (<a href="/products/HA600071" style="font-weight: bold;text-decoration: underline;">HA600071</a>) at 1/2,000 dilution.<br /><br />Lane 1: A431 cell lysate<br />Lane 2: Daudi cell lysate<br />Lane 3: 3T3-L1 cell lysate<br />Lane 4: PC-12 cell lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 65 kDa<br />Observed band size: 77 kDa<br /><br />Exposure time: 2 minutes; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA600071" style="font-weight: bold;text-decoration: underline;">HA600071</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CENPB on different lysates with Mouse anti-CENPB antibody (HA600071) at 1/2,000 dilution.

    Lane 1: A431 cell lysate
    Lane 2: Daudi cell lysate
    Lane 3: 3T3-L1 cell lysate
    Lane 4: PC-12 cell lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 65 kDa
    Observed band size: 77 kDa

    Exposure time: 2 minutes; ECL: K1802;

    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 (HA600071) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of CENPB in SH-SY5Y cells (green). Methanol fixed cells were blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/HA600071" style="font-weight: bold;text-decoration: underline;">HA600071</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of CENPB in SH-SY5Y cells (green). Methanol fixed cells were blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (HA600071, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-CENPB antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA600071" style="font-weight: bold;text-decoration: underline;">HA600071</a>, 1/400) for 30 minutes 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.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-CENPB antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600071, 1/400) for 30 minutes 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.

  • Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-CENPB antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA600071" style="font-weight: bold;text-decoration: underline;">HA600071</a>, 1/100) for 30 minutes 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.

    Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-CENPB antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600071, 1/100) for 30 minutes 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"