Anti-KMT4 / Dot1L antibody [EPR22936-138]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling KMT4 / Dot1L with ab239358 at 1/250 dilution (2.78 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on spermatocytes of human testis. The section was incubated with ab239358 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling KMT4 / Dot1L with ab239358 at 1/4000 dilution (0.17 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on spermatocytes of rat testis. The section was incubated with ab239358 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling KMT4 / Dot1L with ab239358 at 1/4000 dilution (0.17 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on spermatocytes of mouse testis. The section was incubated with ab239358 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins

• WB

Lab

Western blot - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 26199140).

Fresh lysates are recommended on WB test.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-KMT4 / Dot1L antibody [EPR22936-138] (ab239358) at 1/1000 dilution

Lane 1:

HeLa (human cervix 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:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 4:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 5:

HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 6:

NIH/3T3 (mouse embryonic fibroblast), 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

Predicted band size: 185 kDa

Observed band size: 185 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ 293T(Human embryonic kidney epithelial cell) cells and 5 µg of ab239358 [EPR22936-138]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ 293T(Human embryonic kidney epithelial cell) cells and 5 µg of ab239358 [EPR22936-138]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ 293T(Human embryonic kidney epithelial cell) cells and 5 µg of ab239358 [EPR22936-138]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Lab

Western blot - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 26199140).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-KMT4 / Dot1L antibody [EPR22936-138] (ab239358) at 1/1000 dilution

All lanes:

C6 (rat glial tumor glial cell), whole cell lysate at 10 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

Predicted band size: 185 kDa

Observed band size: 185 kDa

false