Anti-KDM4B / JMJD2B antibody [EPR25240-51]

• IHC

Supplier Data

Immunohistochemistry - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue labeling KDM4B / JMJD2B with ab305058 at 1/200 (2.3 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human endometrial carcinoma. The section was incubated with ab305058 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Immunohistochemical analysis of paraffin-embedded cell pellets labeling KDM4B / JMJD2B with ab305058 at 1/200 (2.3 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on (A) parental HAP1 cell pellet, (B) SW480 cell pellet, and (C) HEK-293 cell pellet, no staining on (D) KDM4B knockout HAP1 cell pellet. The section was incubated with ab305058 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling KDM4B / JMJD2B with ab305058 at 1/200 (2.3 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human breast. The section was incubated with ab305058 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Immunohistochemical analysis of paraffin-embedded human pancreatic carcinoma tissue labeling KDM4B / JMJD2B with ab305058 at 1/200 (2.3 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human pancreatic carcinoma. The section was incubated with ab305058 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Blocking/dilution buffer : 5% NFDM/TBST. This antibody does not cross-react with human JMJD2A/KDM4A, JMJD2C/KDM4C or JMJD2D/KDM4D. Exposure time : 10 seconds.

All lanes:

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (ab305058) at 1/1000 dilution

Lane 1:

Flag-tagged human JMJD2A/KDM4A recombinant protein 10ng

Lane 2:

Flag-tagged human JMJD2B/KDM4B recombinant protein 10ng

Lane 3:

Flag-tagged human JMJD2C/KDM4C recombinant protein 10ng

Lane 4:

Flag-tagged human JMJD2D/KDM4D recombinant protein 10ng

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: 160 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Blocking/dilution buffer : 5% NFDM/TBST. In Western blot, ab305058 was shown to bind specifically to KDM4B / JMJD2B. A band was observed at 160 kDa in wild-type HAP1 cell lysate whereas no signal observed at this size in KDM4B / JMJD2B knockout cell line. Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. The bands between 50 kDa and 80 kDa are likely to be degraded target fragments. Exposure time : 158 seconds.

All lanes:

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (ab305058) at 1/1000 dilution

Lane 1:

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

Lane 2:

KDM4B / JMJD2B knockout HAP1 whole cell lysate 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 1/50000 dilution

Observed band size: 160 kDa

false

Exposure time: 158s

• WB

Supplier Data

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Blocking/dilution buffer : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (ab305058) at 1/1000 dilution

All lanes:

SW480 (human colorectal adenocarcinoma epithelial cell) 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: 160 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (AB305058)

Western blot : Anti-KDM4B antibody [EPR25240-51] (ab305058) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab305058 was shown to bind specifically to KDM4B. A band was observed at 135 kDa in wild-type A549 cell lysates with no signal observed at this size in KDM4B knockout cell line. To generate this image, wild-type and KDM4B knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-KDM4B / JMJD2B antibody [EPR25240-51] (ab305058) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

KDM4B knockout A549 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 122 kDa

Observed band size: 135 kDa

false