CST Epigenetics

Understanding the role epigenetics plays in cancer initiation and progression is critical when looking for new methods of prevention, detection, and treatment. We’ve put together a starter’s guide on important epigenetic biomarkers, regulators, and histone modifications for several types of cancer.

Check out the links below to get started.

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Mixed Lineage Leukemia (MLL)

Mixed lineage leukemia is characterized by chromosomal translocations of the mixed lineage leukemia (MLL) gene to form fusion proteins with partner proteins. Many of these MLL-fusion partners belong to the super-elongation complex involved in regulating transcriptional elongation. MLL-fusions can create oncogenic transcription factors that cause aberrant transcriptional regulation of oncogenes or tumor suppressors to initiate oncogenesis.

 

  1. MLL1 is processed into two polypeptides, both which are subunits of the wild-type MLL complex. The amino terminus of MLL1 is what is included in the fusion proteins that cause leukemogenesis. We recommend using a carboxy-terminal specific antibody for detecting wild-type MLL1 and an amino-terminal specific antibody for detecting MLL-fusion proteins.

    – MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) #14197 – WB, IP, IF

MLL1
Western blot analysis of extracts from MLL1 wild-type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.

 
  1. – MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen) #14689 – WB, IP

 

  1. DOT1L interacts with MLL translocation partners in the super-elongation complex and is required for MLL-driven leukemiogenesis. DOT1L is thought to drive gene expression through the methylation of histone H3 at Lys79.

    – DOT1L (D1W4Z) Rabbit mAb #77087 – WB, IP, ChIP

Figure 2
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells and either DOT1L (D1W4Z) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP Human PabPC1 Exon 1 Primers #77410, Human RPLP0 Exon 2 Primers, and SimpleChIP Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

 

 

  1. p300 and CBP are additional MLL-fusion proteins that can drive gene expression through histone acetylation.

    – p300 (D8Z4E) Rabbit mAb #86377 – WB, IP, IF, IHC
    – p300 (D2X6N) Rabbit mAb #54062 – WB, IP, ChIP, ChIP-seq

Figure 3
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and p300 (D2X6N) Rabbit mAb using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using SimpleChIP® ChIP-Seq DNA Library Prep Kit for Illumina® #56795, and sequenced on the Illumina NextSeq. The figure shows binding across FDPS gene.

 
  1. – CBP (D6C5) Rabbit mAb #7389 – WB, IP, IF, ChIP, ChIP-seq

 

  1. Changes in levels of these marks as detected by ChIP/ChIP-seq may indicate altered recruitment of MLL1, DOT1L, and CBP/p300 to target genes.

    – Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751 – WB, IF, F, IHC, ChIP, ChIP-seq
    – Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb #5427 – WB, IF, F, IHC, ChIP
    – Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb #74073 – WB, IP, IF, F, IHC, ChIP
    – Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 – WB, IP, IF, F, IHC, ChIP, ChIP-seq