Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59, rabbit monoclonal

17 kDa

Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Dimethylation of Lys79 on histone H3 is associated with active gene transcription and is a marker for euchromatin. Histone H3 is methylated on Lys79 by the histone methyltransferase Dot1L..

Epitope: Lys79

KLH-conjugated synthetic peptide containing the sequence DF_me2KTD corresponding to dimethyl-lysine at position 79 of histone H3. The immunizing sequence is identical in human, mouse, rat and yeast. A C-terminal cysteine was added to facilitate conjugation.

Anti-dimethyl-Histone H3 (Lys79) Antibody, clone NL59 is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys79) also known as H3K79me2, Histone H3 (di methyl K79) & has been validated in ChIP, WB, DB, Mplex.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 4 µL of Negative Control Supernatant, or 4 µL of Anti-dimethyl-Histone H3 (Lys79) and the Magna ChIP A Kit (Cat. # 17-610). Successful immunoprecipitation of dimethyl-Histone H3 (Lys79) associated DNA fragments was verified by qPCR using Control Primers
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Chromatin Immunoprecipitation: A representative lot of this antibody has been shown by an independent laboratory to work in ChIP.

ChIP-Seq Analysis:
A representative lot of this antibody was used by an independent laboratory for ChIP-Seq. See Egelhofer, T.A., et al. (2011). See Easwaran, H., et al. (2012).See Suzuki, H., et al. (2011).

Dot Blotting: Specificity of a representative lot confirmed by the ability of a 1:2500 dilution of the antibody to recognize peptides corresponding to regions of histone H3 with various modifications

Dot Blot Analysis: Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 04-835-S, Anti-dimethyl Histone H3 (Lys79) Antibody, clone NL59 (1:500 dilution). Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Histones

Broad species cross-reactivity expected due to sequence homology

Recognizes dimethyl-histone H3 (Lys79).

Culture supernantant in 0.05% sodium azide

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

routinely evaluated by immunoblot of acid extracted proteins from HeLa cells

Replaces: 05-835

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