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Western blot - Histone H3 antibody - ChIP Grade (ab1791)
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ChIP - Histone H3 antibody - ChIP Grade (ab1791)
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ChIP - Histone H3 antibody - ChIP Grade (ab1791)
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Product Name
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Histone H3 antibody - ChIP Grade
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See all Histone H3 - unmodified antibodies (20)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Histone H3 - ChIP Grade
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Immunogen
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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.)
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Reacts with
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Hu, Ms, Rat, At, Ce, Chk, Dog, Dm, InMtj, N cra, Sc, S pom, Tx gg, Try c, Xl
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Predicted to react
(species key)
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Many, Mamm
(Due to sequence homology)
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Tested applications
(see key)
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CHIPseq, ChIP, ChIP/Chip, Dot, Flow Cyt, ICC/IF, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
CHIPseq: Use at an assay dependent dilution.
ChIP: Use 2µg for 106 cells.
ChIP on chip: Use at an assay dependent dilution.
Dot: 1/10,000.
Flow Cyt: Use at an assay dependent dilution.
ICC: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent dilution.
IF: 1/500.
IHC-P: 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP: Use at an assay dependent dilution.
WB: 1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). Can be blocked with Histone H3 peptide (ab12149).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Chromatin and Nuclear Signaling >> Histones >> H3 >> Unmodified
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Relevance
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Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fibre is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.
Covalent modifications of the canonical core histones, including acetylation, phosphorylation, methylation, and monoubiquitination are used to mark nucleosomes to create chromatin domains with a range of functions. The information encoded by histone modifications can contribute to the formation and/or maintenance of transcriptionally active and inactive chromatin in response to various signalling pathways.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.50 - 1.00 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Histone H3 antibody - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Histone H3 antibody - ChIP Grade images, references, products related to ab1791 and other tools.
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Histone H3 antibody - ChIP Grade images:
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 Western blot - Histone H3 antibody - ChIP Grade (ab1791)
All lanes : Histone H3 antibody - ChIP Grade (ab1791) at 1/1000 dilution
Lane 1 : A431 Whole Cell Lysate
Lane 2 : Jurkat Whole Cell Lysate
Lane 3 : 293 Whole Cell Lysate
Lane 4 : A431 Whole Cell Lysate with Histone H3 peptide (ab12149) at 1 µg/ml
Lane 5 : Jurkat Whole Cell Lysate with Histone H3 peptide (ab12149) at 1 µg/ml
Lane 6 : 293 Whole Cell Lysate with Histone H3 peptide (ab12149) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Exposure time : 10 seconds
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ChIP - Histone H3 antibody - ChIP Grade (ab1791)
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.
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ChIP - Histone H3 antibody - ChIP Grade (ab1791)
Hela cells cultured on coverslips were fixed with 4% paraformaldehyde and then stained with ab1791 (green) at a working dilution of 1/500. The DNA stained with DAPI is shown in red. (100x magnification). Kirk McManus, University of British Columbia
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ChIP - Histone H3 antibody - ChIP Grade (ab1791)
Chromatin from Xenopus laevis oocytes was prepared according to the Abcam X-ChIP protocol. Oocytes were fixed with formaldehyde for 10 min. The ChIP was performed with 25 mg of chromatin, 3 mg of ab7834 (anti-H3, light blue) and 3 µg of ab1791 (anti-H3, dark blue), and 20 ml of Protein A/G sepharose beads. A non-specific antibody was used as a control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).
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Flow Cytometry - Histone H3 antibody - ChIP Grade (ab1791)
ab1791 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cell colonies were trypsinized and incubated with the antibody 1ug/1.5 x 105cells in a permeabilization buffer. A PE conjugated goat anti-rabbit antibody was used as the secondary. This image is courtesy of an Abreview submitted by Prof Albrecht Müller
See Abreview
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody - ChIP Grade (ab1791)
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. This image is courtesy of an anonymous Abreview
See Abreview
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Western blot - Histone H3 antibody - ChIP Grade (ab1791)
All lanes : Histone H3 antibody - ChIP Grade (ab1791) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 3 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12Hr)
Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate
Lane 5 : S.pombe Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
ab1791 is tested in western blot on a range of species. We recommend loading higher amounts of protein (20-30ug) to increase the signal in yeast lysates
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Immunocytochemistry/ Immunofluorescence - Histone H3 antibody - ChIP Grade (ab1791)
ICC/IF image of ab1791 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1791, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 5% PFA fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HeLa and Hek293 cells at 1µg/ml.
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Western blot - Histone H3 antibody - ChIP Grade (ab1791)
Predicted band size : 15 kDa
John E. Mueller and J. Ruth German (Mary Bryk lab)
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References for Histone H3 antibody - ChIP Grade (ab1791)
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This product has been used in: (two most recent references)
Mehedint MG et al. Choline deficiency alters global histone methylation and epigenetic marking at the Re1 site of the calbindin 1 gene. FASEB J 24:184-95 (2010). WB; Mouse. PubMed: 19752176
Kumar SV & Wigge PA H2A.Z-Containing Nucleosomes Mediate the Thermosensory Response in Arabidopsis. Cell 140:136-147 (2010). ChIP; Arabidopsis thaliana. PubMed: 20079334
See all 247 publication references for this product.
If you publish research using ab1791 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Histone H3
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