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CHD4 Recombinant Rabbit Monoclonal Antibody [JA33-40]

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Specification

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Catalog# ET1704-53

CHD4 Recombinant Rabbit Monoclonal Antibody [JA33-40]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CHD4 Recombinant Rabbit Monoclonal Antibody [JA33-40]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human CHD4 aa 1,811-1,860.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 218 kDa

Positive Control

293T cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, 293T, human breast carcinoma tissue, human colon carcinoma tissue.

Conjugation

unconjugated

Clone Number

JA33-40

RRID

AB_3070500

Product Features

Form

Liquid

Concentration

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), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:400

  • FC

  • 1:50-1:100

Target

Function

In the intact cell, DNA closely associates with histones and other nuclear proteins to form chromatin. The remodeling of chromatin is believed to be a critical component of transcriptional regulation and a major source of this remodeling is brought about by the acetylation of nucleosomal histones. Acetylation of lysine residues in the amino terminal tail domain of histone results in an allosteric change in the nucleosomal conformation and an increased accessibility to transcription factors by DNA. Conversely, the deacetylation of histones is associated with transcriptional silencing. Chromatin structure alteration may be brought about by the action of ATP-dependent multiprotein complexes. One such complex is the mSin3 corepressor complex, which contains mSin3, the histone deacetylases HDAC1 and HDAC2, the associated proteins SAP 30 and SAP 18, and the autoantigens Mi2-α and Mi2-β.

Background References

1. D\'Alesio C et al. RNAi screens identify CHD4 as an essential gene in breast cancer growth.Oncotarget 7:80901-80915 (2016).

2. Petell CJ et al. An epigenetic switch regulates de novo DNA methylation at a subset of pluripotency gene enhancers during embryonic stem cell differentiation. Nucleic Acids Res 44:7605-17 (2016).

Sequence Similarity

Belongs to the SNF2/RAD54 helicase family.

Subcellular Location

Nucleus. Cytoplasm.

UNIPROT

SwissProt: Q14839 Human

SwissProt: Q6PDQ2 Mouse

Synonyms

ATP dependent helicase CHD4 antibody

ATP-dependent helicase CHD4 antibody

CHD 4 antibody

CHD-4 antibody

CHD4 antibody

CHD4_HUMAN antibody

Chromodomain helicase DNA binding protein 4 antibody

Chromodomain-helicase-DNA-binding protein 4 antibody

Mi 2 autoantigen 218 kDa protein antibody

Mi 2b antibody

Expand

Images

  • Western blot analysis of CHD4 on different lysates with Rabbit anti-CHD4 antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>) at 1/500 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: NIH/3T3 cell lysate<br />Lane 3: C2C12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 218 kDa<br />Observed band size: 260 kDa<br /><br />Exposure time: 30 seconds;<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/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>) at 1/500 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CHD4 on different lysates with Rabbit anti-CHD4 antibody (ET1704-53) at 1/500 dilution.

    Lane 1: 293T cell lysate
    Lane 2: NIH/3T3 cell lysate
    Lane 3: C2C12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 218 kDa
    Observed band size: 260 kDa

    Exposure time: 30 seconds;

    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 (ET1704-53) at 1/500 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of CHD4 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of CHD4 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-53, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-CHD4 antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>) at 1/400 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>) at 1/400 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.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-CHD4 antibody (ET1704-53) at 1/400 dilution.

    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 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-53) at 1/400 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.

  • Flow cytometric analysis of CHD4 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of CHD4 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-53, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-CHD4 antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>) at 1/400 dilution.<br /><br />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 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-53" style="font-weight: bold;text-decoration: underline;">ET1704-53</a>) at 1/400 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.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-CHD4 antibody (ET1704-53) at 1/400 dilution.

    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 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-53) at 1/400 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.

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

Citation