Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146]

• ChIP-seq

Lab

ChIP-sequencing - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326104 [EPR29602-146]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326104 [EPR29602-146]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Lab

Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

KDM4A / JHDM3A / JMJD2A was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab326104 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326104 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab326104 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326104 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 180 seconds.

To minimize protein degradation, cells (lanes 5-8) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (ab326104) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab326104 at 1/30 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab326104 in HeLa whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 150 kDa

false

Exposure time: 180s

• ChIP-seq

Lab

ChIP-sequencing - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326104 [EPR29602-146]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Chromatin was prepared from 3T3-L1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326104 [EPR29602-146]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Chromatin was prepared from 3T3-L1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326104 [EPR29602-146]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Chromatin was prepared from 3T3-L1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326104 [EPR29602-146]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Lab

Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

KDM4A / JHDM3A / JMJD2A was immunoprecipitated from 0.35 mg 3T3-L1(mouse embryonic fibroblast) whole cell lysate with ab326104 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326104 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 3T3-L1(mouse embryonic fibroblast) whole cell lysate
Lane 2 : ab326104 IP in 3T3-L1(mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326104 in 3T3-L1 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 180 seconds.

To minimize protein degradation, cells (lanes 5-8) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (ab326104) at 1/1000 dilution

Lane 1:

3T3-L1(mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

ab326104 at 1/30 IP in 3T3-L1(mouse embryonic fibroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab326104 in 3T3-L1 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 150 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells (lanes 5-8) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-5 : 59 seconds; Lanes 6-8 : 180 seconds

All lanes:

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (ab326104) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

HeLa (human cervical adenocarcinoma epithelial cell) fresh whole cell lysate at 20 µg

Lane 7:

293T (human embryonic kidney epithelial cell) fresh whole cell lysate at 20 µg

Lane 8:

NIH/3T3 (mouse embryonic fibroblast) fresh whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 150 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab326104 was shown to bind specifically to KDM4A / JHDM3A / JMJD2A. Target of interest was observed at 150 kDa in wild-type HPA1 cell lysates (lane 1) with no signal observed at this size in KDM4A / JHDM3A / JMJD2A knockout cell line.

To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (ab326104) at 1/1000 dilution

Lane 1:

Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

KDM4A / JHDM3A / JMJD2A knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (AB326104)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with recombinant human JMJD2B / KDM4B, KDM4C / GASC1 / JMJD2C, KDM4B / JMJD2B by western blot.

In Western blot, Anti-DDDDK-tag antibody (1 : 5000) (150kDa, 65kDa ).

All lanes:

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR29602-146] (ab326104) at 1/1000 dilution

Lane 1:

DDDDK-tagged human KDM4A / JHDM3A / JMJD2A recombinant protein at 10 ng

Lane 2:

DDDDK-tagged human JMJD2B / KDM4B recombinant protein at 10 ng

Lane 3:

DDDDK-tagged human KDM4C / GASC1 / JMJD2C recombinant protein at 10 ng

Lane 4:

DDDDK-tagged human KDM4B / JMJD2B recombinant protein at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 150 kDa,65 kDa

false

Exposure time: 59s