DOT1L antibody (pAb)

RRID: AB_2793406
Aliases: DOT1, KMT4
Catalog No: 39953 Format: 100 µl ¥86,000 Buy
Catalog No: 39954 Format: 10 µl ¥24,000 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Affinity Purified
Host:
Rabbit
Molecular Weight:
200 kDa
Reactivity:
Human

Applications

Western Blot Validated

Application Notes

Applications Validated by Active Motif:
WB*: 1:500 - 1:1,000 dilution

The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection by Western blot. Individual optimization may be required.

*Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot.

Immunogen

This DOT1L antibody was raised against a peptide derived from the C-terminal region of human DOT1L.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 

DOT1L antibody (pAb) tested by Western blot.
Nuclear extract of HEK293 cells (40 µg per lane) probed with DOT1L antibody (pAb) at a dilution of 1:500.

Background

DOT1L (Disruptor of Telomeric silencing 1-like, also known as KMT4) is the mammalian homologue of the yeast Dot1 protein and the Drosophila Grappa protein. DOTL1 is the enzyme responsible for lysine 79 methylation of histone H3. DOT1L can promote an oncogenic pattern of gene expression through binding with several MLL fusion partners found in acute leukemia, thus leading to aberrant methylation of lysine 79 at MLL target genes. In contrast to most other histone lysine methyltransferases, DOT1L does not contain a SET domain. Biochemical studies suggest that recombinant DOT1L is active on recombinant nucleosomes.

Positive Control

Active Motif's 293 nuclear extract (Catalog No. 36033) can be used as a positive control for DOT1L antibody.

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting