MLL1/KMT2A

MLL1/KMT2A CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2004
Pack Size: 100 µL
  • Type:  Monoclonal [BL-175-7E8]
  • Host:  Rabbit
  • Target Size:  431 kDa
  • Format:  Purified IgG
  • Reactivity:  Human
  • Applications: CUT&RUN, WB, IP

Description

This antibody meets EpiCypher’s “CUTANA Compatible” criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping. Every lot of a CUTANA Compatible antibody is tested in the indicated CUTANA approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. MLL1 antibody produces CUT&RUN peaks above background (Figure 1) that overlap with H3K4me3 (Figures 1-2), consistent with its known role as the catalytic subunit of the MLL1/MLL histone methyltransferase complex.

Validation Data

Figure 1: MLL1 enrichment at annotated transcription start sites (TSSs) in CUT&RUN
CUT&RUN was performed using 500,000 K562 cells with MLL1 and control antibodies (0.5 µg each; IgG negative control, EpiCypher 13-0042; H3K4me3 positive control, EpiCypher 13-0041). Sequencing reads were aligned to annotated TSSs (+/- 2 kbp) of 18,793 genes. High, medium, and low signal is ranked by intensity (top to bottom) and reflected by red, yellow, and blue colors, respectively. All rows aligned relative to MLL1.

Figure 2: MLL1 CUT&RUN peaks and functional overlap
Two representative gene loci from the CUT&RUN data in Figure 1 are shown. MLL1 peaks overlap with H3K4me3 CUT&RUN peaks (EpiCypher 13-0041), consistent with the reported function of MLL1 as an H3K4 histone methyltransferase. Images were generated using the Integrative Genomics Viewer (Broad Institute).

Figure 3: Western blot detection of human MLL1
Whole cell lysates were isolated from HEK293T, HeLa, MCF-7, HEP-G2, A-549, and SW620 cells using NETN lysis buffer. Fifty micrograms (50 µg) of lysate were loaded onto SDS-PAGE gel and analyzed under standard western blot conditions using MLL1 antibody (1:1,000 dilution).

Figure 4: Immunoprecipitation of human MLL1
EpiCypher MLL1 antibody (10 µL) was used to immunoprecipitate whole cell lysates isolated from HEK293T cells using NETN lysis buffer (0.5 mg per IP). A negative control IgG antibody and positive control antibodies to various MLL1 epitopes (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto SDS-PAGE gel (20% of IP loaded) and probed via western blot with EpiCypher MLL1 antibody (1:1,000 dilution).

Technical Information

Immunogen
A synthetic peptide corresponding to human MLL1 amino acids 720 to 780.
Formulation
Purified recombinant monoclonal antibody (1.0 mg/mL) in borate buffered saline (BBS) pH 8.2 with 0.09% sodium azide.
Storage and Stability
Stable for 1 year at 4°C from date of receipt.

Application Notes

Recommended Dilutions:

CUT&RUN: 0.5 µg

Western Blot (WB): 1:1,000

Immunoprecipitation (IP): 10 µL / 0.5 mg lysate

Documents & Resources

Additional Info

This product is provided for commercial sale under license from Bethyl Laboratories, Inc.

Applications Key:

ChIP: Chromatin immunoprecipitation
CUT&RUN: Cleavage Under Targets and Release Using Nuclease
CUT&Tag: Cleavage Under Targets and Tagmentation
E: ELISA
FACS: Flow cytometry
ICC: Immunocytochemistry
IF: Immunofluorescence
IHC: Immunohistochemistry
IP: Immunoprecipitation
L: Luminex
WB: Western Blot

Reactivity Key:

B: Bovine
Ce: C. elegans
Ch: Chicken
Dm: Drosophila
Eu: Eukaryote
H: Human
M: Mouse
Ma: Mammal
R: Rat
Sc: S. cerevisiae
Sp: S. pombe
WR: Wide Range (predicted)
X: Xenopus
Z: Zebrafish
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