Anti-Histone H3抗体
参阅全部 Histone H3 一抗
兔多克隆抗体to Histone H3
Rabbit
ab18521 recognises unmodified and modified forms of histone H3
适用于: IP, ICC/IF, WB, IHC-Pmore details
与反应: Cow, Human, Saccharomyces cerevisiae
预测可用于: Mouse, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster
Synthetic peptide corresponding to Human Histone H3 aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
Database link: P68431
(Peptide available as ab14949)
ICC/IF: HeLa cells. WB: Calf Thymus Histone Preparation Nuclear Lysate. HeLa Histone Preparation Nuclear Lysate - Colcemid-treated. Histone H3.1 Recombinant Protein. Whole cell lysate prepared from Saccharomyces cerevisiae WT (BY4741, delta hht1 HHT2 and delta hht1 hh2-K9, 14, 18R). IHC-P: Human testis tissue. IP: HeLa whole cell extract.
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.
浓度
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab18521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 |
|---|---|---|
| IP | Use a concentration of 5 µg/ml. | |
| ICC/IF | (2) | Use a concentration of 1 - 5 µg/ml. |
| WB | (5) | Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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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: 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
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
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
H3 histone family member E pseudogene antibody
H3 histone family, member A antibody
H3/A antibody
Western blot - Anti-Histone H3 antibody (ab18521)
All lanes : Anti-Histone H3 antibody (ab18521) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 2 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated at 2.5 µg
Lane 3 : Histone H2A Recombinant Protein at 0.1 µg
Lane 4 : Histone H3.1 Recombinant Protein at 0.1 µg
Lane 5 : Histone H4 Recombinant Protein at 0.1 µg
Secondary
All lanes : 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 kDawhy is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody (ab18521)
IHC image of Histone H3 staining in human testis 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 ab18521, 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.
Immunoprecipitation - Anti-Histone H3 antibody (ab18521)
Histone H3 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18521.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 17kDa; Histone H3
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody (ab18521)
ab18521 staining Histone H3 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 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 ab18521 at 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 100% methanol (5 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 H3 antibody (ab18521)This image was kindly supplied by Dr Tobias Eisenberg by Abreview
All lanes : Anti-Histone H3 antibody (ab18521) at 1/250 dilution
Lane 1 : Whole cell lysate prepared from Saccharomyces cerevisiae WT (BY4741).
Lane 2 : Whole cell lysate prepared from Saccharomyces cerevisiae delta hht1 HHT2.
Lane 3 : Whole cell lysate prepared from Saccharomyces cerevisiae delta hht1 hh2-K9, 14, 18R.
Lysates/proteins at 500000 cells per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
Exposure time: 10 minutes
Blocking performed using 3% BSA for 16 hours at 4°C.
Lysates prepared using 5% Trichloroethane.
Western blot - Anti-Histone H3 antibody (ab18521)
All lanes : Anti-Histone H3 antibody (ab31602) at 1 µg/ml
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with Human Histone H3 peptide (ab14949) at 1 µg/ml
Lane 3 : Calf thymus histone lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml
Lane 4 : Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/ml
Lane 5 : Calf thymus histone lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 15 kDa
Observed band size: 15 kDa
ab18521 recognises histone H3 as well as modified histone H3 at approximately 15 kDa(lane1). This is demonstrated by the blocking using the immunising unmodified histone H3 peptide (lane2), histone H3 mono methyl K4 (lane3), histone H3 di methyl K4 (lane4) and histone H3 tri methyl K4 (lane5).
