Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade
| Name | Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab2886 |
| Prices | $403.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | ChIP ICC/IF IP Immunodiffusion ICC/IF ICC/IF WB Array |
| Species Reactivities | Mouse, Rat, Human, Yeast, Broad species, Mammalian |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Histone H3, mono methylated at K79 |
| Blocking Peptide | Human Histone H3 (mono methyl K79) peptide |
| Description | Rabbit Polyclonal |
| Gene | HIST3H3 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ab2886 staining Histone H3 (mono methyl K79) in human differentiated haematopoietic stem cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/800) for 12 hours at 4°C. An undiluted Alexa Fluor® 488-conjugated goat polyclonal anti-rabbit IgG was used as the secondary antibody.Gating Strategy: Isotype negative control (white).See Abreview
Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µgSecondaryHuman NFIB / NF1B2 peptide (ab95051) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
All batches of ab2886 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - mono methyl K79 peptide (ab4555), indicating that this antibody specifically recognises the Histone H3 - mono methyl K79 modification.ab4558 - Histone H3 - unmodifiedab1771 - Histone H3 - di methyl K9ab4555 - Histone H3 - mono methyl K79ab4556 - Histone H3 - di methyl K79ab4557 - Histone H3 - tri methyl K79ab4560 - Histone H4 - di methyl K79
This image was kindly supplied as part of the review submitted by Dr Geza Fejes-Toth.
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab2886 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR. Primers and probes are located in the first kb of the transcribed region.
All lanes : Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886) at 1/300 dilutionLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 3 : Testis (Rat) Tissue Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Chromatin was prepared from whole cell lysate of normal rat liver and liver cancer cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. 5 µg of the primary antibody was used in 1/100 dilution and it was incubated with the sample for 16 hours at 4°C in a commercially available ChIP dilution buffer. The immunoprecipitated DNA was quantified by real time PCR. ChIP results show that the Histone H3 (mono methyl K79) and GAPDH genes are expressed in higher levels in liver cancer cells than in normal liver cells.See Abreview
Histone H3 (mono methyl K79) was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H3 (mono methyl K79) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2886.Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.Band: 18kDa: Histone H3 (mono methyl K79).
Product References
Histone H3 K79 methylation states play distinct roles in UV-induced sister - Histone H3 K79 methylation states play distinct roles in UV-induced sister
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Allele-specific H3K79 Di- versus trimethylation distinguishes opposite parental - Allele-specific H3K79 Di- versus trimethylation distinguishes opposite parental
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The Dot1 histone methyltransferase and the Rad9 checkpoint adaptor contribute to - The Dot1 histone methyltransferase and the Rad9 checkpoint adaptor contribute to
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The histone H3K79 methyltransferase Dot1L is essential for mammalian development - The histone H3K79 methyltransferase Dot1L is essential for mammalian development
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