Anti-Histone H2A抗体- ChIP Grade
参阅全部 Histone H2A 一抗
兔多克隆抗体to Histone H2A - ChIP Grade
Rabbit
适用于: ICC/IF, WB, IP, ChIPmore details
与反应: Mouse, Cow, Human
Synthetic peptide within Human Histone H2A aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as ab19751)
WB: HeLa nuclear extract. Calf thymus histone preparation. Histone H2A Recombinant Protein. ChIP: Chromatin from U-2 OS cells. IP: Histone H2A IP in HeLa whole cell 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.
浓度
批次浓度范围 100 µg 浓度为 0.5 - 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab18255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 | (4) | 1/200. see Abreview submitted by Kirk McManus |
| WB | (13) | Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). |
| IP | Use a concentration of 5 µg/ml. | |
| ChIP | (2) | Use at an assay dependent concentration. Every new batch of this antibody is tested at Abcam in ChIP |
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SwissProt: P04908 Human
SwissProt: P0C0S8 Human
SwissProt: P20671 Human
SwissProt: Q16777 Human
SwissProt: Q6FI13 Human
SwissProt: Q7L7L0 Human
SwissProt: Q8IUE6 Human
SwissProt: Q93077 Human
SwissProt: Q96KK5 Human
SwissProt: Q96QV6 Human
SwissProt: Q99878 Human
SwissProt: P10812 Mouse
SwissProt: P22752 Mouse
Unigene: 121017 Human
Unigene: 248174 Human
Entrez Gene: 3012 Human
Entrez Gene: 317772 Human
Entrez Gene: 8335 Human
Entrez Gene: 8338 Human
Entrez Gene: 319166 Mouse
Omim: 142720 Human
Omim: 602786 Human
Omim: 602797 Human
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ChIP - Anti-Histone H2A antibody - ChIP Grade (ab18255)Reamon-Buettner and Borlak PLoS One. 2012;7(6):e38531. doi: 10.1371/journal.pone.0038531. Epub 2012 Jun 6. Fig 4. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Myc affects the incorporation levels of histone variant H2A.Z.
qChIP was performed using specific antibodies recognizing A. H2A.Z, B. H2A.Zac, Panel D. H2A, E. H2AK5ac and G. H1. All the qChIP values are expressed as % of input and normalized for total histone H3, with the exception of C and F, where H2A.Z acetylation is noramlized for H2A.Z density, and H2AK5 acetylation is normalized for H2A density, respectively. The box plots show the fold change distribution of each acetylated residue for the two subpopulations.
Western blot - Anti-Histone H2A antibody - ChIP Grade (ab18255)
All lanes : Anti-Histone H2A antibody - ChIP Grade (ab18255) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 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
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
Exposure time: 3 minutes
Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A antibody - ChIP Grade (ab18255)Colpitts et al PLoS One. 2011;6(9):e24365. doi: 10.1371/journal.pone.0024365. Epub 2011 Sep 1. Fig 2.
DENV C colocalizes with histones in Huh7 liver cells.
DEN2 C colocalized with H2A (A), H2B (B), H3 (C) and H4 (D) in Huh7 cells. Cells were transfected with GFP-DEN2 C and fixed in 4% paraformaldehyde 48 h post-transfection. Cells were stained with antibodies against histones and a TRITC secondary antibody. Cells were counterstained with DAPI to visualize the nucleus. GFP-DEN2 C expression is green, histone staining is red and DAPI is blue.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A antibody - ChIP Grade (ab18255)
ICC/IF image of ab18255 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in 100% methanol (5 min) then permeabilized using 0.1% PBS-Triton and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab18255 at 1 µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.
Immunoprecipitation - Anti-Histone H2A antibody - ChIP Grade (ab18255)
Histone H2A - ChIP Grade was immunoprecipitated using 0.5 mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extract, 5 µg of Rabbit polyclonal to and 50 µL of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10 min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 min under agitation.
Proteins were eluted by addition of 40 µL SDS loading buffer and incubated for 10 min at 70°C; 10 µL of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18255.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 14kDa, non specific bands - 42kDa: We are unsure as to the identity of this extra band; Histone H2A - ChIP Grade
ChIP - Anti-Histone H2A antibody - ChIP Grade (ab18255)
Chromatin was prepared from U-2 OS (Human bone osteosarcoma epithelial cell line) 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, 6 µL of ab18255 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
Western blot - Anti-Histone H2A antibody - ChIP Grade (ab18255)Colpitts et al PLoS One. 2011;6(9):e24365. doi: 10.1371/journal.pone.0024365. Epub 2011 Sep 1. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
DENV expression disrupts histone oligomerization.
Panel A and B shown:
Huh7 cells were transfected with DEN2 C and/or infected with DEN2 24 h post-transfection. Cells were lysed 24 h post-infection (48 h post-transfection) and lysates were run on 4–12% SDS-PAGE gel. Gels were used in a Western blotting assay with antibodies against histones H2A ab18255 (A), H2B (B), H3 (C) and H4 (D); monomers, dimers and octamers are indicated. Gels were stripped and reprobed with an antibody against actin as a protein loading control. The same amount of protein was loaded in each lane for each gel as a control for expression.
Western blot - Anti-Histone H2A antibody - ChIP Grade (ab18255)
All lanes : Anti-Histone H2A antibody - ChIP Grade (ab18255) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate at 20 µg
Lane 2 : HeLa nuclear lysate at 20 µg
Lane 3 : Calf thymus histone lysate at 20 µg
Lane 4 : HeLa lysate at 1 µg/ml with Human Histone H2A peptide (ab19751) at 1 µg/ml
Lane 5 : HeLa nuclear lysate at 1 µg/ml with Human Histone H2A peptide (ab19751) at 1 µg/ml
Lane 6 : Calf thymus histone lysate at 1 µg/ml with Human Histone H2A peptide (ab19751) at 1 µg/ml
Predicted band size: 14 kDa
Observed band size: 14 kDa
Additional bands at: 22 kDa (possible cross reactivity)
ab18255 is partially blocked by the immunizing peptide ab19751. There is an additional band at 22kDa in HeLa lysate which is attributed to cross-reactivity.
Western blot - Anti-Histone H2A antibody - ChIP Grade (ab18255)This image is courtesy of an Abreview submitted by Ragnhild Eskeland
All lanes : Anti-Histone H2A antibody - ChIP Grade (ab18255) at 1/1000 dilution
Lane 1 : Native recombinant octamers K562 cells
Lane 2 : Recombinant Human octamers containing H2A
Lane 3 : Recombinant Human octamers containing H2A.Z.2.1
Lane 4 : Recombinant Human octamers containing H2A.Z.1
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 15 kDawhy is the actual band size different from the predicted?
Exposure time: 5 minutes
