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HDAC9 Recombinant Rabbit Monoclonal Antibody [JU30-44]

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Specification

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Catalog# ET1706-36

HDAC9 Recombinant Rabbit Monoclonal Antibody [JU30-44]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

HDAC9 Recombinant Rabbit Monoclonal Antibody [JU30-44]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human HDAC9 aa 1-50 / 1,011.

Species Reactivity

Human

Validated Applications

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

Molecular Weight

Predicted band size: 111 kDa

Positive Control

RD cell lysate, K-562 cell lysate, K-562, A431, human brain tissue, human bladder cancer tissue.

Conjugation

unconjugated

Clone Number

JU30-44

RRID

AB_3070657

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • 1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:500-1:2,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

In the intact cell, DNA closely associates with histones and other nuclear proteins to form chromatin. The remodeling of chromatin is a critical component of transcriptional regulation and the acetylation of nucleosomal histones is a major source of this remodeling. 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. Several mammalian proteins function as nuclear histone acetylases, including GCN5, PCAF (p300/CBP-associated factor), p300/CBP, HAT1 and the TFIID subunit TAF II p250. Conversely, the deacetylation of histones is associated with transcriptional silencing. The histone deacetylases (HDAC) include HDAC1-9. HDAC9 and HDAC9a are two alternatively spliced isoforms of HDAC9. HDAC9a is 132 amino acids shorter than HDAC9, but both isoforms contain the HDAC catalytic domain, remain capable of deacetylase activity and repress myoctye enhancer-binding factor 2-mediated transcription.

Background References

1. Di Giorgio E et al. The co-existence of transcriptional activator and transcriptional repressor MEF2 complexes influences tumor aggressiveness. PLoS Genet 13:e1006752 (2017).

2. Bourgeois CT et al. HDAC9 is an epigenetic repressor of kidney angiotensinogen establishing a sex difference. Biol Sex Differ 8:18 (2017).

Sequence Similarity

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

Tissue Specificity

Broadly expressed, with highest levels in brain, heart, muscle and testis. Isoform 3 is present in human bladder carcinoma cells (at protein level).

Post-translational Modification

Phosphorylated on Ser-220 and Ser-450; which promotes 14-3-3-binding, impairs interaction with MEF2, and antagonizes antimyogenic activity. Phosphorylated on Ser-240; which impairs nuclear accumulation (By similarity). Isoform 7 is phosphorylated on Tyr-1010. Phosphorylated by the PKC kinases PKN1 and PKN2, impairing nuclear import.; Sumoylated.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q9UKV0 Human

Synonyms

HD 7 antibody

HD 7B antibody

HD 9 antibody

HD7 antibody

HD7B antibody

HD9 antibody

HDAC 7 antibody

HDAC 7B antibody

HDAC 9 antibody

HDAC 9B antibody

Expand

Images

  • Western blot analysis of HDAC9 on different lysates with Rabbit anti-HDAC9 antibody (<a href="/products/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/2,000 dilution.<br /><br />Lane 1: RD cell lysate<br />Lane 2: K-562 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 111 kDa<br />Observed band size: 160 kDa<br /><br />Exposure time: 1 minute 18 seconds; 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/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/2,000 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.

    Western blot analysis of HDAC9 on different lysates with Rabbit anti-HDAC9 antibody (ET1706-36) at 1/2,000 dilution.

    Lane 1: RD cell lysate
    Lane 2: K-562 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 111 kDa
    Observed band size: 160 kDa

    Exposure time: 1 minute 18 seconds; 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 (ET1706-36) at 1/2,000 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of HDAC9 on different lysates with Rabbit anti-HDAC9 antibody (<a href="/products/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-HDAC9 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 111 kDa<br />Observed band size: 160 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/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 HDAC9 on different lysates with Rabbit anti-HDAC9 antibody (ET1706-36) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-HDAC9 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 111 kDa
    Observed band size: 160 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 (ET1706-36) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of K-562 cells labeling HDAC9 with Rabbit anti-HDAC9 antibody (<a href="/products/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</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-HDAC9 antibody (<a href="/products/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</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.

    Immunocytochemistry analysis of K-562 cells labeling HDAC9 with Rabbit anti-HDAC9 antibody (ET1706-36) 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-HDAC9 antibody (ET1706-36) 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.

  • ICC staining of HDAC9 in A431 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/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of HDAC9 in A431 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 (ET1706-36, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit 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 brain tissue with Rabbit anti-HDAC9 antibody (<a href="/products/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/2,000 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/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/2,000 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 brain tissue with Rabbit anti-HDAC9 antibody (ET1706-36) at 1/2,000 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 (ET1706-36) at 1/2,000 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 bladder cancer tissue with Rabbit anti-HDAC9 antibody (<a href="/products/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/500 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/ET1706-36" style="font-weight: bold;text-decoration: underline;">ET1706-36</a>) at 1/500 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 bladder cancer tissue with Rabbit anti-HDAC9 antibody (ET1706-36) at 1/500 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 (ET1706-36) at 1/500 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.

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Citation