Anti-KMT4 / Dot1L antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT4 / Dot1L antibody (AB72454)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling KMT4 / Dot1L with ab72454 at 1/200 (1µg/ml). Detection : DAB.

• IP

Unknown

Immunoprecipitation - Anti-KMT4 / Dot1L antibody (AB72454)

HeLa whole cell lysate ( 1 mg for IP, 20% of IP loaded) with ab72454 at 3 µg/mg lysate for IP, followed by ab72454 at 1 µg/ml for WB.
Detection : Chemiluminescence with an exposure time of 10 seconds.

All lanes:

Immunoprecipitation - Anti-KMT4 / Dot1L antibody (ab72454)

Predicted band size: 185 kDa

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• WB

Unknown

Western blot - Anti-KMT4 / Dot1L antibody (AB72454)

All lanes:

Western blot - Anti-KMT4 / Dot1L antibody (ab72454) at 0.04 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

HeLa whole cell lysate at 15 µg

Lane 3:

HeLa whole cell lysate at 5 µg

Predicted band size: 185 kDa

Observed band size: 100 kDa,185 kDa

true

Exposure time: 3min

• WB

CiteAb

Western blot - Anti-KMT4 / Dot1L antibody (AB72454)

KMT4 / Dot1L western blot using anti-KMT4 / Dot1L antibody ab72454. Publication image and figure legend from Liu, D., Zhang, X. X., et al., 2018, Nat Commun, PubMed 29712898.

ab72454 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab72454 please see the product overview.

C/EBPβ recruits the methyltransferase DOT1L to target genes that methylate H3K79. a Venn diagrams showing the overlap of C/EBPβ- and DOT1L-targeted genes (chi-squared test). b Correlation analysis of C/EBPβ binding sites and DOT1L binding sites. Red dots represent genes with two peak centers located less than 2500 bp apart. c Representative results of C/EBPβ and DOT1L ChIP-qPCR. Primers were chosen according to the ChIP-seq results (upper panel). d C/EBPβ interacts with DOT1L. Lysates from C13* cells were immunoprecipitated (IP) with a mouse anti-C/EBPβ antibody and analyzed by western blot using a rabbit anti-DOT1L antibody (upper panel) or the reciprocal (lower panel). e ChIP-reChIP experiments with anti-C/EBPβ and anti-DOT1L antibodies. Mouse IgG (mIgG) and rabbit IgG (rIgG) were used as negative controls. f Meta-analysis of the averaged DOT1L ChIP-seq signal of genes across a ±10 kb genomic region flanking the TSS. g Normalized C/EBPβ and DOT1L ChIP-seq signal of the representative C/EBPβ-DOT1L co-targeted genes (EREG and SLC38A1). h Analysis of DOT1L ChIP-qPCR (green), H3K79me2/me3 ChIP-qPCR (blue), and RT-qPCR (purple) in C13* cells. Gene names in red indicate documented cisplatin-resistance genes in ovarian cancer. i The protein levels of C/EBPβ-DOT1L co-targeted genes in the indicated OV2008 cells were detected by western blot. Uncropped images of blots are shown in Supplementary Figure 25. *p < 0.05; ***p < 0.001

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