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Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human Histone H3.
(Peptide available as ab12149.)
Our Abpromise guarantee covers the use of ab1791 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|CHIPseq||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.|
|IHC-P||1/100 - 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Electron Microscopy||1/50. Customer feedback|
|ICC/IF||Use a concentration of 1 µg/ml.|
|ChIP||Use 2µg for 106 cells.|
|IP||Use at an assay dependent concentration.|
|WB||1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).|
|ChIP/Chip||Use at an assay dependent concentration.|
|IHC - Wholemount||Use at an assay dependent concentration. PubMed: 22219645|
|ICC||1/100 - 1/500.|
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 ab1791 (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 (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
ab1791 staining rat testes tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval prior to blocking with 5% serum for 30 minutes at 22°C. The primary antibody was diluted 1/400 and incubated with the sample for 30 minutes at 22°C. A HRP-conjugated goat anti-rabbit antibody diluted 1/400 was used as the secondary.
Image courtesy of Dr Roman Hudec by Abreview.
Histone H3 immunogold detection in HeLa cells.
Ulltra-thin sections of paraformaldehyde-fixed, Lowicryl K4M embedded cells were incubated sequentially with ab1791 antibody and an anti-rabbit antibody coupled to 10 nm gold particles. Notice absence of chromatin within Interchromatin Granules (IG), a reservoir of spliceosomal snRNPs, and high labeling of patches of condensed chromatin close to the nuclear envelope (arrows). Nu: nucleus, Cyt: cytoplasm.
Sample : human
Type : HeLa cells
Fixative : either paraformaldehyde 4% or 1.6% glutaraldehyde in 0.1M Millonig’s buffer.
Embedding : in Lowicryl 4KM at -20°C under UV.
Ultrathin-sections deposited on formvar-coated carbonated EM-grids.
Blocking step: 5% BSA for 30 seconds at RT.
Primary antibody: Ab1791 diluted 1/50 in PBS, for 1h at RT.
Secondary antibody: BBI International anti-rabbit antibody coupled to 10 nm gold particles
Additional comments: A highly specific antibody working similarly on thin-sections of glutaraldehyde or paraformaldehyde-fixed cells. Ideal for testing antigeneicity, for detecting micronuclei, chromatin content of nuclear organelles or bodies…
John E. Mueller and J. Ruth German (Mary Bryk lab)
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"