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KDM4A Recombinant Rabbit Monoclonal Antibody [PSH03-67]

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Specification

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Catalog# HA722026

KDM4A Recombinant Rabbit Monoclonal Antibody [PSH03-67]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

KDM4A Recombinant Rabbit Monoclonal Antibody [PSH03-67]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human KDM4A aa 351-650 / 1,064.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Target Molecular Weight

Predicted band size: 121 kDa

Positive Control

A549 cell lysate, HEK-293 cell lysate, HeLa cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, NCCIT, NIH/3T3, human colon tissue, mouse brain tissue, rat colon tissue.

Conjugation

unconjugated

Clone Number

PSH03-67

RRID

AB_3096617

Product Features

Form

Liquid

Concentration

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

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:500

  • FC

  • 1:1,000

Target

Function

Lysine-specific demethylase 4A is an enzyme that in humans is encoded by the KDM4A gene. This gene is a member of the Jumonji domain 2 (JMJD2) family and encodes a protein with a JmjN domain, a JmjC domain, a JD2H domain, two TUDOR domains, and two PHD-type zinc fingers. This nuclear protein belongs to the alpha-ketoglutarate-dependent hydroxylase superfamily. It functions as a trimethylation-specific demethylase, converting specific trimethylated histone on histone H3 lysine 9 and 36 residues to the dimethylated form and lysine 9 dimethylated residues to monomethyl, and as a transcriptional repressor. Alterations in this gene have been found associated with chromosomal instability that leads to cancer.

Background References

1. Zhang W et al. Targeting KDM4A epigenetically activates tumor-cell-intrinsic immunity by inducing DNA replication stress. Mol Cell. 2021 May

2. Young NL et al. Mechanistic insights into KDM4A driven genomic instability. Biochem Soc Trans. 2021 Feb

Subcellular Location

Nucleus.

UNIPROT

SwissProt: O75164 Human

SwissProt: Q8BW72 Mouse

Entrez Gene: 313539 Rat

Synonyms

JHDM3A antibody

JmjC domain containing histone demethylation protein 3A antibody

JmjC domain-containing histone demethylation protein 3A antibody

JMJD2 antibody

JMJD2A antibody

jumonji C domain containing histone demethylase 3A antibody

Jumonji domain containing 2 antibody

Jumonji domain containing 2A antibody

Jumonji domain containing protein 2A antibody

Jumonji domain-containing protein 2A antibody

Expand

Images

  • Western blot analysis of KDM4A on different lysates with Rabbit anti-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549 cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: HepG2 cell lysate<br />Lane 5: NIH/3T3 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 136 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 121 kDa<br />Observed band size: 150 kDa

    Western blot analysis of KDM4A on different lysates with Rabbit anti-KDM4A antibody (HA722026) at 1/2,000 dilution.

    Lane 1: A549 cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: HepG2 cell lysate
    Lane 5: NIH/3T3 cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 136 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA722026, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 121 kDa
    Observed band size: 150 kDa

  • Immunocytochemistry analysis of NCCIT cells labeling KDM4A with Rabbit anti-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</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 NCCIT cells labeling KDM4A with Rabbit anti-KDM4A antibody (HA722026) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-KDM4A antibody (HA722026) 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 (M1305-2, 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling KDM4A with Rabbit anti-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</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 NIH/3T3 cells labeling KDM4A with Rabbit anti-KDM4A antibody (HA722026) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-KDM4A antibody (HA722026) 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 (M1305-2, 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.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/1,000 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/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/1,000 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 colon tissue with Rabbit anti-KDM4A antibody (HA722026) at 1/1,000 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 (HA722026) at 1/1,000 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 mouse brain tissue with Rabbit anti-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/1,000 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/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/1,000 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 mouse brain tissue with Rabbit anti-KDM4A antibody (HA722026) at 1/1,000 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 (HA722026) at 1/1,000 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 rat colon tissue with Rabbit anti-KDM4A antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/1,000 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/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>) at 1/1,000 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 rat colon tissue with Rabbit anti-KDM4A antibody (HA722026) at 1/1,000 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 (HA722026) at 1/1,000 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.

  • Flow cytometric analysis of NCCIT cells labeling KDM4A.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722026" style="font-weight: bold;text-decoration: underline;">HA722026</a>, 1μg/mL) (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 NCCIT cells labeling KDM4A.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722026, 1μg/mL) (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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