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HDAC4 Recombinant Rabbit Monoclonal Antibody [SD08-35]

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Specification

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Catalog# ET1612-51

HDAC4 Recombinant Rabbit Monoclonal Antibody [SD08-35]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

HDAC4 Recombinant Rabbit Monoclonal Antibody [SD08-35]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human HDAC4 aa 11-125/1084.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 119 kDa

Positive Control

HCT 116 cell lysate, human colon cancer tissue, human testis tissue, HeLa.

Conjugation

unconjugated

Clone Number

SD08-35

RRID

AB_3070125

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

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100-1:500

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:200

  • FC

  • 1:1,000

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. Several mammalian proteins have been identified as nuclear histone acetylases, including GCN5, p300/CBP, PCAF (p300/CBPassociated factor), HAT1, and the TFIID subunit TAF II p250. Mammalian HDAC1 (also designated HD1), HDAC2 (also designated RPD3) and HDAC3-6, have been identified as histone deacetylases.

Background References

1. Yang Y et al. The interaction of the atm genotype with inflammation and oxidative stress. PLoS One 9:e85863 (2014).

2. Sharma A et al. Calcium-mediated histone modifications regulate alternative splicing in cardiomyocytes. Proc Natl Acad Sci U S A 111:E4920-8 (2014).

Sequence Similarity

Belongs to the histone deacetylase family. HD type 2 subfamily.

Tissue Specificity

Ubiquitous.

Post-translational Modification

Phosphorylated by CaMK4 at Ser-246, Ser-467 and Ser-632. Phosphorylation at other residues by CaMK2D is required for the interaction with 14-3-3. Phosphorylation at Ser-350, within the PxLPxI/L motif, impairs the binding of ANKRA2 but generates a high-affinity docking site for 14-3-3.; Sumoylation on Lys-559 is promoted by the E3 SUMO-protein ligase RANBP2, and prevented by phosphorylation by CaMK4.

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: P56524 Human

Synonyms

AHO3 antibody

BDMR antibody

EC 3.5.1.98 antibody

HA6116 antibody

HD 4 antibody

HD4 antibody

HDAC 4 antibody

HDAC A antibody

HDAC4 antibody

HDAC4_HUMAN antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of HDAC4 on different lysates with Rabbit anti-HDAC4 antibody (<a href="/products/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/1,000 dilution.<br /><br />Lane 1: HCT 116-si NT cell lysate<br />Lane 2: HCT 116-si HDAC4 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 119 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<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/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/1,000 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.

    ☑ Knockdown (KD)

    Western blot analysis of HDAC4 on different lysates with Rabbit anti-HDAC4 antibody (ET1612-51) at 1/1,000 dilution.

    Lane 1: HCT 116-si NT cell lysate
    Lane 2: HCT 116-si HDAC4 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 119 kDa
    Observed band size: 150 kDa

    Exposure time: 3 minutes; ECL: K1801;

    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 (ET1612-51) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-HDAC4 antibody (<a href="/products/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/200 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/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/200 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 cancer tissue with Rabbit anti-HDAC4 antibody (ET1612-51) at 1/200 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 (ET1612-51) at 1/200 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 testis tissue with Rabbit anti-HDAC4 antibody (<a href="/products/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/200 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/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/200 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 testis tissue with Rabbit anti-HDAC4 antibody (ET1612-51) at 1/200 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 (ET1612-51) at 1/200 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.

  • Immunocytochemistry analysis of HeLa cells labeling HDAC4 with Rabbit anti-HDAC4 antibody (<a href="/products/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HDAC4 antibody (<a href="/products/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling HDAC4 with Rabbit anti-HDAC4 antibody (ET1612-51) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HDAC4 antibody (ET1612-51) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HeLa cells labeling HDAC4.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1612-51" style="font-weight: bold;text-decoration: underline;">ET1612-51</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling HDAC4.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-51, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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