Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal

• ChIP-seq

Lab

ChIP-sequencing - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (AB319035)

Chromatin was prepared from ES-D3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab319035 [EPR27970-7]. 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-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (AB319035)

Chromatin was prepared from ES-D3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab319035 [EPR27970-7]. 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-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (AB319035)

Chromatin was prepared from ES-D3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab319035 [EPR27970-7]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

The Input control is also shown.

• WB

Supplier Data

Western blot - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (AB319035)

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

Negative control : HCT 116.

The identity of the lower MW band at approximately 60 kDa is unknown.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-3 : 180 seconds; Lane 4 : 15 seconds; Lanes 5-12 : 103 seconds

All lanes:

Western blot - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (ab319035) at 1/1000 dilution

Lane 1:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

ES-D3 D3  (mouse blastocyst-derived embryonic stem cell) whole cell lysate at 20 µg

Lane 5:

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

Lane 6:

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

Lane 7:

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

Lane 8:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 9:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 10:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 11:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 12:

SH-SY5Y (human neuroblastoma 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: 80 kDa,36 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (AB319035)

Dot blot analysis of KMT2B / MLL4 - C-terminal using ab319035 at 1 : 1000 (0.512 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : His-tagged human KMT2B / MLL4 fragment
Lane2 : His-tagged human KMT2A / MLL fragment

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

This antibody does not cross-react with human KMT2A / MLL.
In dot blot, anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (ab319035) at 1/1000 dilution

Lane 1:

His-tagged human KMT2B / MLL4 fragment

Lane 2:

His-tagged human KMT2A / MLL fragment

Secondary

All lanes:

Dot 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

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (AB319035)

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

Low expression : liver (PMID : 10409430).

The identity of the lower MW band at approximately 37 kDa in Lane 1 is unknown.

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-KMT2B / MLL4 antibody [EPR27970-7] - C-terminal (ab319035) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Rat liver tissue lysate at 20 µg

Lane 4:

Rat testis tissue 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: 80 kDa,36 kDa

false

Exposure time: 180s