Anti-Histone H3 (acetyl K27)抗体- ChIP Grade
参阅全部 Histone H3 一抗
兔多克隆抗体to Histone H3 (acetyl K27) - ChIP Grade
Rabbit
From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.
适用于: ICC/IF, WB, IHC-P, ChIP, PepArrmore details
与反应: Mouse, Rat, Cow, Human, Recombinant fragment
预测可用于: Chicken, Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Monkey, Zebrafish, Plasmodium falciparum, Rice, Cyanidioschyzon merolae
Synthetic peptide corresponding to Human Histone H3 aa 1-100 (acetyl K27) conjugated to keyhole limpet haemocyanin.
(Peptide available as ab24404)
WB : HeLa (human cervix adenocarcinoma epithelial cell) cell lysate - Sodium butyrate-treated, HeLa (human cervix adenocarcinoma epithelial cell) nuclear lysate (triton enriched), NIH/3T3 (mouse embryonic fibroblast cell line) nuclear lysate (triton enriched) and PC-12 (rat adrenal gland pheochromocytoma cell) nuclear lysate (triton enriched). ICC/IF: HeLa cells
Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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ab4729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 |
|---|---|---|
| ICC/IF | (23) | Use a concentration of 0.5 µg/ml. Can be used with paraformaldehyde- or methanol- fixed cells. |
| WB | (20) | Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody. |
| IHC-P | (4) | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
| ChIP | (27) | Use 2 µg for 25 µg of chromatin. We recommend GAPDH positive control ChIP primer pair ab267832 as a positive control. |
| PepArr | Use a concentration of 0.2 - 0.02 µg/ml. |
Entrez Gene: 8350 Human
Entrez Gene: 8351 Human
Entrez Gene: 8352 Human
Entrez Gene: 8353 Human
Entrez Gene: 8354 Human
Entrez Gene: 8355 Human
Entrez Gene: 8356 Human
Entrez Gene: 8357 Human
Entrez Gene: 8358 Human
Entrez Gene: 8968 Human
Entrez Gene: 319152 Mouse
Entrez Gene: 319153 Mouse
Entrez Gene: 360198 Mouse
Entrez Gene: 97908 Mouse
Omim: 602810 Human
SwissProt: P84229 Chicken
SwissProt: P68431 Human
SwissProt: P68433 Mouse
Unigene: 132854 Human
Unigene: 247813 Human
Unigene: 247814 Human
Unigene: 248176 Human
Unigene: 443021 Human
Unigene: 484990 Human
Unigene: 532144 Human
Unigene: 533292 Human
Unigene: 546315 Human
Unigene: 586261 Human
Unigene: 591778 Human
Unigene: 221301 Mouse
Unigene: 261657 Mouse
Unigene: 377874 Mouse
Unigene: 390558 Mouse
Unigene: 397328 Mouse
Unigene: 138090 Rat
H3 histone family member E pseudogene antibody
H3 histone family, member A antibody
H3/A antibody
H31_HUMAN antibody
H3F3 antibody
H3FA antibody
Hist1h3a antibody
HIST1H3B antibody
HIST1H3C antibody
HIST1H3D antibody
HIST1H3E antibody
HIST1H3F antibody
HIST1H3G antibody
HIST1H3H antibody
HIST1H3I antibody
HIST1H3J antibody
HIST3H3 antibody
histone 1, H3a antibody
Histone cluster 1, H3a antibody
Histone H3 3 pseudogene antibody
Histone H3.1 antibody
Histone H3/a antibody
Histone H3/b antibody
Histone H3/c antibody
Histone H3/d antibody
Histone H3/f antibody
Histone H3/h antibody
Histone H3/i antibody
Histone H3/j antibody
Histone H3/k antibody
Histone H3/l antibody
ChIP - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)This image is courtesy of an anonymous Abreview
ChIP was performed with human thyroid cancer cell lysate and 1 µg/µg of ab4729. Lysates were incubated with the primary antibody for 16 hours at 4°C. positive control promoter: GAPDH promoter; positive control enhancer: enhancer region of a published target (10.1093/nar/gkx802); negative control: published negative region (10.1093/nar/gkx802).
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
ab4729 staining Histone H3 (acetyl K27) 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 ab4729 at 1µg/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 magenta). 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.
ChIP - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 2 µg of ab4729 (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 H3 (acetyl K27) antibody - ChIP Grade (ab4729)
IHC image of ab4729 staining Histone H3 (acetyl K27) in human colon formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH 6 for 20 minutes. The section was then incubated with ab4729, 5 µg/ml, for 15 minutes 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.
No primary antibody was used in the negative control (shown on the inset).
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
Peptide Array - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
All batches of ab4729 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 - acetyl K27 peptide (ab24404), indicating that this antibody specifically recognises the Histone H3 - acetyl K27 modification.
ab24404 - Histone H3 - acetyl K27
ab15591 - Histone H3 - acetyl K14
ab24003 - Histone H3 - acetyl K18
ab17163 - Histone H3 unmodified
ab48359 - Histone H3 - acetyl K23
ab41409 - Histone H3 - acetyl K36
ab15662 - Histone H4 - acetyl K12
ab16635 - Histone H3 acetyl K9
Western blot - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
All lanes : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 1 µg/ml
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) cell lysate - Sodium butyrate-treated
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) nuclear lysate (triton enriched)
Lane 3 : NIH/3T3 (mouse embryonic fibroblast cell line) nuclear lysate (triton enriched)
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma cell) nuclear lysate (triton enriched)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
Blocking buffer : 2% BSA block
Gel type : MES
Western blot - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
All lanes : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 1/2500 dilution
Lane 1 : Untreated Mouse MEF cell lysate
Lane 2 : 0.4 µM Trichostatin A treatment for 18 hr Mouse MEF cell lysate
Lysates/proteins at 9 µg per lane.
Secondary
All lanes : Donkey Anti-Rabbit IgG H&L (HRP) (ab6802) at 1/20000 dilution
Predicted band size: 15 kDa
Western blot - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
Lanes 1 & 3 : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 0.2 µg/ml
Lanes 2 & 4 : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 0.1 µg/ml
Lanes 1-2 : Calf thymus histone lysate
Lanes 3-4 : Calf thymus histone lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 2 µg
Lysates/proteins at 1 µg per lane.
Secondary
All lanes : Goat anti-rabbit (HRP) at 1/2000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
ab4729 specifically recognises acetyl K27 histone H3 in catlf thymus histone lysate, which is specifically blocked using the immunizing peptide ab24404.
Western blot - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were incubated at 37°C for 6 hours with vehicle control (0 μM) and different concentrations of sodium butyrate (ab120948). Increased expression of histone H3 (acetyl K27)(ab4729) in HeLa cells correlates with an increase in sodium butyrate concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 2.5 μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab4927 at 1 μg/ml and ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10,000 dilution and visualised using ECL development solution.
Western blot - Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)
Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 1 µg/ml + HeLa (Human epithelial cell line from cervix adenocarcinoma) histone preparation, nuclear Lysate - Butyrate treated at 2.5 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 seconds
