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KDM4B Recombinant Rabbit Monoclonal Antibody [JE36-58]

Usd: 385

Specification

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Catalog# HA721350

KDM4B Recombinant Rabbit Monoclonal Antibody [JE36-58]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

KDM4B Recombinant Rabbit Monoclonal Antibody [JE36-58]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human KDM4B aa 97-146 / 1,096.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 122 kDa

Positive Control

HeLa cell lysate, HCT116 cell lysate, SW480 cell lysate, mouse testis tissue lysate, rat testis tissue lysates, human breast carcinoma tissue, HeLa, SW480.

Conjugation

unconjugated

Clone Number

JE36-58

RRID

AB_3072467

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

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200

  • IF-Cell

  • 1:250

Target

Function

Lysine-specific demethylase 4B is an enzyme that in humans is encoded by the KDM4B gene. KDM4B belongs to the alpha-ketoglutarate-dependent hydroxylase superfamily. Histone demethylase that specifically demethylates 'Lys-9' of histone H3, thereby playing a role in histone code. Does not demethylate histone H3 'Lys-4', H3 'Lys-27', H3 'Lys-36' nor H4 'Lys-20'. Only able to demethylate trimethylated H3 'Lys-9', with a weaker activity than KDM4A, KDM4C and KDM4D. Demethylation of Lys residue generates formaldehyde and succinate. Plays a critical role in the development of the central nervous system (CNS).

Background References

1. Deng P et al. Loss of KDM4B exacerbates bone-fat imbalance and mesenchymal stromal cell exhaustion in skeletal aging. Cell Stem Cell. 2021 Jun

2. Duncan AR et al. Heterozygous Variants in KDM4B Lead to Global Developmental Delay and Neuroanatomical Defects. Am J Hum Genet. 2020 Dec

Subcellular Location

Nucleus.

UNIPROT

SwissProt: O94953 Human

SwissProt: Q91VY5 Mouse

Synonyms

JHDM3B antibody

JmjC domain-containing histone demethylation protein 3B antibody

JMJD2B antibody

Jumonji domain containing 2B antibody

Jumonji domain containing protein 2B antibody

Jumonji domain-containing protein 2B antibody

Kdm4b antibody

KDM4B_HUMAN antibody

KIAA0876 antibody

Lysine (K) specific demethylase 4B antibody

Expand

Images

  • Western blot analysis of KDM4B on different lysates with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HCT116 cell lysate (20 µg/Lane)<br />Lane 3: SW480 cell lysate (20 µg/Lane)<br />Lane 4: Mouse testis tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 121 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 42 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/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of KDM4B on different lysates with Rabbit anti-KDM4B antibody (HA721350) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HCT116 cell lysate (20 µg/Lane)
    Lane 3: SW480 cell lysate (20 µg/Lane)
    Lane 4: Mouse testis tissue lysate (40 µg/Lane)

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

    Exposure time: 42 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 (HA721350) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of KDM4B on rat testis tissue lysates with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 122 kDa<br />Observed band size: 140 kDa<br /><br />Exposure time: 1 minute;<br /><br />6% 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/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of KDM4B on rat testis tissue lysates with Rabbit anti-KDM4B antibody (HA721350) at 1/1,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 122 kDa
    Observed band size: 140 kDa

    Exposure time: 1 minute;

    6% 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 (HA721350) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</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/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</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 breast carcinoma tissue with Rabbit anti-KDM4B antibody (HA721350) 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 (HA721350) 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 KDM4B with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/250 dilution in 2% negative goat serum 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. 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 HeLa cells labeling KDM4B with Rabbit anti-KDM4B antibody (HA721350) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KDM4B antibody (HA721350) at 1/250 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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 SW480 cells labeling KDM4B with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KDM4B antibody (<a href="/products/HA721350" style="font-weight: bold;text-decoration: underline;">HA721350</a>) at 1/250 dilution in 2% negative goat serum 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. 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 SW480 cells labeling KDM4B with Rabbit anti-KDM4B antibody (HA721350) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KDM4B antibody (HA721350) at 1/250 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.

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