Anti-KMT2D / MLL2 antibody [EPR29598-577]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT2D / MLL2 antibody [EPR29598-577] (AB324486)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling KMT2D / MLL2 with ab324486 at 1/1000 (0.508 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human stomach. The primary antibody was incubated 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 a 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-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT2D / MLL2 antibody [EPR29598-577] (AB324486)

Immunohistochemical analysis of paraffin-embedded A : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet. B : KMT2D knockout HAP1 cell pellet tissue labeling KMT2D / MLL2 with ab324486 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on A : Wild-type HAP1 cell pellet, no staining on B : KMT2D knockout HAP1 cell pellet. The primary antibody was incubated 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 a 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-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT2D / MLL2 antibody [EPR29598-577] (AB324486)

Immunohistochemical analysis of paraffin-embedded Human diffuse large B-cell lymphoma tissue labeling KMT2D / MLL2 with ab324486 at 1/1000 (0.508 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human diffuse large B-cell lymphoma (PMID : 26366712). The primary antibody was incubated 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 a 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

• IP

Supplier Data

Immunoprecipitation - Anti-KMT2D / MLL2 antibody [EPR29598-577] (AB324486)

KMT2D / MLL2 was immunoprecipitated from 0.35 mg Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate with ab324486 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324486 at 1/1000 dilution.

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied for Immunoprecipitation as soon as possible.

All lanes:

Immunoprecipitation - Anti-KMT2D / MLL2 antibody [EPR29598-577] (ab324486) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 10 µg

Lane 2:

ab324486 IP in Wild-type HAP1 (human chronic myelogenous leukemia near-haploid 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 ab324486 in Wild-type HAP1 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: 593 kDa

false

Exposure time: 76s

• WB

Supplier Data

Western blot - Anti-KMT2D / MLL2 antibody [EPR29598-577] (AB324486)

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

In Western blot, ab324486 was shown to bind specifically to KMT2D. Target of interest was observed at 593 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in KMT2D knockout cell line (lane 2).

To minimize protein degradation, cells 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-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-KMT2D / MLL2 antibody [EPR29598-577] (ab324486) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 40 µg

Lane 2:

KMT2D knockout HAP1 whole cell lysate at 40 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: 593 kDa,124 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-KMT2D / MLL2 antibody [EPR29598-577] (AB324486)

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

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

Exposure time : Lane 1 : 26 seconds; Lane 2 : 37 seconds; Lane 3 : 92 seconds; Lane 4 : 103 seconds.

All lanes:

Western blot - Anti-KMT2D / MLL2 antibody [EPR29598-577] (ab324486) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast 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: 593 kDa

false