Anti-Histone H3 (di methyl K27) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab24684
• Prices: $403.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: WB IP IHC-P ICC/IF ICC/IF ChIP
• Species Reactivities: Mouse, Bovine, Human, Yeast, C. elegans, Plant
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, di methylated at K27
• Blocking Peptide: Human Histone H3 (di methyl K27) peptide
• Description: Rabbit Polyclonal
• Gene: HIST3H3
• Conjugate: Unconjugated

Product images

All lanes : Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684) at 1 µg/mlLane 1 : Calf thymus histone lysateLane 2 : Calf thymus histone lysate with Human Histone H3 peptide (ab17163) at 0.5 µg/mlLane 3 : Calf thymus histone lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/mlLane 4 : Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/mlLane 5 : Calf thymus histone lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/mlLane 6 : Calf thymus histone lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/mlLane 7 : Calf thymus histone lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/mlLane 8 : Calf thymus histone lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/mlLane 9 : Calf thymus histone lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/mlLane 10 : Calf thymus histone lysate with Human Histone H3 (di methy

Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of  ab24684 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

ab24684 staining Histone H3 (di methyl K27) 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 (1/600) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat polyclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.Gating Strategy: Isotype negative control (white).See Abreview

ICC/IF image of ab24684 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab24684, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

IHC image of Histone H3 (di methyl K27) staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab24684, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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Stragier E, Massart R, Salery M, Hamon M, Geny D, Martin V, Boulle F, Lanfumey L. Mol Psychiatry. 2015 Mar;20(3):405-12.

Comparative analysis of genome-wide chromosomal histone modification patterns in - Comparative analysis of genome-wide chromosomal histone modification patterns in

He S, Yan S, Wang P, Zhu W, Wang X, Shen Y, Shao K, Xin H, Li S, Li L. PLoS One. 2014 May 12;9(5):e97364.

Inhibition of DNA methylation alters chromatin organization, nuclear positioning - Inhibition of DNA methylation alters chromatin organization, nuclear positioning

Bockor VV, Barisic D, Horvat T, Maglica Z, Vojta A, Zoldos V. PLoS One. 2014 Aug 5;9(8):e103954.

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Heinemann B, Nielsen JM, Hudlebusch HR, Lees MJ, Larsen DV, Boesen T, Labelle M, Gerlach LO, Birk P, Helin K. Nature. 2014 Oct 2;514(7520):E1-2.

Reduced H3K27me3 and DNA hypomethylation are major drivers of gene expression in - Reduced H3K27me3 and DNA hypomethylation are major drivers of gene expression in

Bender S, Tang Y, Lindroth AM, Hovestadt V, Jones DT, Kool M, Zapatka M, Northcott PA, Sturm D, Wang W, Radlwimmer B, Hojfeldt JW, Truffaux N, Castel D, Schubert S, Ryzhova M, Seker-Cin H, Gronych J, Johann PD, Stark S, Meyer J, Milde T, Schuhmann M, Ebinger M, Monoranu CM, Ponnuswami A, Chen S, Jones C, Witt O, Collins VP, von Deimling A, Jabado N, Puget S, Grill J, Helin K, Korshunov A, Lichter P, Monje M, Plass C, Cho YJ, Pfister SM. Cancer Cell. 2013 Nov 11;24(5):660-72.

Epigenetics of eu- and heterochromatin in inverted and conventional nuclei from - Epigenetics of eu- and heterochromatin in inverted and conventional nuclei from

Eberhart A, Feodorova Y, Song C, Wanner G, Kiseleva E, Furukawa T, Kimura H, Schotta G, Leonhardt H, Joffe B, Solovei I. Chromosome Res. 2013 Aug;21(5):535-54.

Sen1p contributes to genomic integrity by regulating expression of ribonucleotide - Sen1p contributes to genomic integrity by regulating expression of ribonucleotide

Golla U, Singh V, Azad GK, Singh P, Verma N, Mandal P, Chauhan S, Tomar RS. PLoS One. 2013 May 31;8(5):e64798.

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Mysliwiec MR, Carlson CD, Tietjen J, Hung H, Ansari AZ, Lee Y. J Biol Chem. 2012 Jan 6;287(2):1235-41.

Epigenetic reprogramming reverses the relapse-specific gene expression signature - Epigenetic reprogramming reverses the relapse-specific gene expression signature

Bhatla T, Wang J, Morrison DJ, Raetz EA, Burke MJ, Brown P, Carroll WL. Blood. 2012 May 31;119(22):5201-10.