Anti-Histone H4 (acetyl K8)抗体- ChIP Grade
参阅全部 Histone H4 一抗
兔多克隆抗体to Histone H4 (acetyl K8) - ChIP Grade
Rabbit
适用于: IHC-P, ChIP, WB, ICC/IFmore details
与反应: Cow, Human
预测可用于: Mouse, Chicken, Saccharomyces cerevisiae
Synthetic peptide corresponding to Human Histone H4 aa 1-100 (internal sequence) (acetyl K8) conjugated to Keyhole Limpet Haemocyanin (KLH).
Database link: P62805
(Peptide available as ab15824)
ChIP: Chromatin was prepared from U2OS cells. IHC-P: Human breast adenocarcinoma tissue. WB: Calf Thymus Histone Preparation Nuclear Lysate. ICC/IF: HeLa cells.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab15823于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ChIP | (2) | Use 2µl for 106 cells. |
| WB | (1) | Use a concentration of 1 µg/ml. Detects a band of approximately 12 kDa (predicted molecular weight: 11.2 kDa). |
| ICC/IF | (2) | Use a concentration of 0.1 - 1 µg/ml. |
Entrez Gene: 121504 Human
Entrez Gene: 554313 Human
Entrez Gene: 8294 Human
Entrez Gene: 8359 Human
Entrez Gene: 8360 Human
Entrez Gene: 8361 Human
Entrez Gene: 8362 Human
Entrez Gene: 8363 Human
Entrez Gene: 8364 Human
Entrez Gene: 8365 Human
Entrez Gene: 8366 Human
Entrez Gene: 8367 Human
Entrez Gene: 8368 Human
Entrez Gene: 8370 Human
Entrez Gene: 100041230 Mouse
Entrez Gene: 319155 Mouse
Entrez Gene: 319156 Mouse
Entrez Gene: 319157 Mouse
Entrez Gene: 319158 Mouse
Entrez Gene: 319159 Mouse
Entrez Gene: 319160 Mouse
Entrez Gene: 319161 Mouse
Entrez Gene: 320332 Mouse
Entrez Gene: 326619 Mouse
Entrez Gene: 326620 Mouse
Entrez Gene: 674678 Mouse
Entrez Gene: 69386 Mouse
Entrez Gene: 97122 Mouse
Omim: 142750 Human
Omim: 602822 Human
Omim: 602823 Human
Omim: 602824 Human
Omim: 602825 Human
Omim: 602826 Human
Omim: 602827 Human
Omim: 602828 Human
Omim: 602829 Human
Omim: 602830 Human
Omim: 602831 Human
Omim: 602833 Human
SwissProt: P62805 Human
SwissProt: P62806 Mouse
SwissProt: P02309 Saccharomyces cerevisiae
Unigene: 143080 Human
Unigene: 247816 Human
Unigene: 248172 Human
Unigene: 248178 Human
Unigene: 248179 Human
Unigene: 278483 Human
Unigene: 46423 Human
Unigene: 528055 Human
Unigene: 533295 Human
Unigene: 55468 Human
Unigene: 591790 Human
Unigene: 655235 Human
Unigene: 662174 Human
Unigene: 706635 Human
Unigene: 718935 Human
Unigene: 144300 Mouse
Unigene: 14775 Mouse
Unigene: 158272 Mouse
Unigene: 227295 Mouse
Unigene: 228709 Mouse
Unigene: 246720 Mouse
Unigene: 255646 Mouse
Unigene: 260530 Mouse
Unigene: 261642 Mouse
Unigene: 261662 Mouse
Unigene: 261664 Mouse
Unigene: 377875 Mouse
Unigene: 442307 Mouse
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H4/I antibody
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HIST1H4A antibody
HIST1H4B antibody
HIST1H4C antibody
HIST1H4D antibody
HIST1H4E antibody
HIST1H4F antibody
HIST1H4H antibody
HIST1H4I antibody
HIST1H4J antibody
HIST1H4K antibody
HIST1H4L antibody
HIST2H4 antibody
HIST2H4A antibody
Hist4h4 antibody
Histone 1 H4a antibody
Histone 1 H4b antibody
Histone 1 H4c antibody
Histone 1 H4d antibody
Histone 1 H4e antibody
Histone 1 H4f antibody
Histone 1 H4h antibody
Histone 1 H4i antibody
Histone 1 H4j antibody
Histone 1 H4k antibody
Histone 1 H4l antibody
Histone 2 H4a antibody
histone 4 H4 antibody
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MGC24116 antibody
ChIP - Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)
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 ab15823 (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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)
IHC image of histone H4 (acetyl K8) antibody staining in a section of formalin-fixed paraffin-embedded human breast adenocarcinoma* performed on a Leica BONDTM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15823, 0.5ug/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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)
All lanes : Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 peptide (ab15825) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 (acetyl K8) peptide (ab15824) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Histone H4 peptide - acetyl K5 at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 (acetyl K12) peptide (ab15662) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H2A (acetyl K9) peptide (ab54016) at 0.5 µg/ml
Lysates/proteins at 0.5 µg/ml per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 11.2 kDa
Observed band size: 13 kDawhy is the actual band size different from the predicted?
Exposure time: 3 minutes
Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)
ab15823 staining Histone H4 (Acetyl K8) in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab15823 at 0.1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Western blot - Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823)
All lanes : Anti-Histone H4 (acetyl K8) antibody - ChIP Grade (ab15823) at 1 µg/ml
Lane 1 : Sodium butyrate treated hela histone
Lane 2 : Untreated hela histone
Lane 3 : Sodium butyrate treated hela histone with Human Histone H4 (acetyl K8) peptide (ab15824) at 1 µg/ml
Lane 4 : Untreated hela histone with Human Histone H4 (acetyl K8) peptide (ab15824) at 1 µg/ml
Lane 5 : Sodium butyrate treated hela histone with Human Histone H4 peptide (ab15825) at 1 µg/ml
Lane 6 : Untreated hela histone with Human Histone H4 peptide (ab15825) at 1 µg/ml
Lane 7 : Sodium butyrate treated hela histone with Histone H4 peptide - acetyl K5 (ab15596) at 1 µg/ml
Lane 8 : Untreated hela histone with Histone H4 peptide - acetyl K5 (ab15596) at 1 µg/ml
Lane 9 : Sodium butyrate treated hela histone with Human Histone H4 (acetyl K12) peptide (ab15662) at 1 µg/ml
Lane 10 : Untreated hela histone with Human Histone H4 (acetyl K12) peptide (ab15662) at 1 µg/ml
Lane 11 : Sodium butyrate treated hela histone with Histone H2A peptide - acetyl K9 at 1 µg/ml
Lane 12 : Untreated hela histone with Histone H2A peptide - acetyl K9 at 1 µg/ml
Lysates/proteins at 2.5 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 11.2 kDa
Observed band size: 12 kDawhy is the actual band size different from the predicted?
Additional bands at: 22 kDa (possible cross reactivity)
The blot shows that a H4 peptide containing an acetylated K8 residue was able to block recognition of histones by ab15823. Peptides containing other acetylated K residues of H4 and H2A did not block binding of histones by ab15823. This suggests ab15823 specifically recognises Histone H4 acetyl K8.
