Anti-Histone H3 (mono methyl K9)抗体- ChIP Grade
参阅全部 Histone H3 一抗
兔多克隆抗体to Histone H3 (mono methyl K9) - ChIP Grade
Rabbit
Specific for mono-methyl lysine 9 of Histone H3. There is no cross-reactivity with lysine 27 of Histone H3.
适用于: WB, ChIP, ICC/IFmore details
与反应: Cow, Human
预测可用于: Caenorhabditis elegans, a wide range of other species, Mammals
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab1771)
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: 1% BSA, 98.98% PBS
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 0.9 - 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab8896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 |
|---|---|---|
| WB | (5) | Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). |
| ChIP | (2) | Use 2 µg for 25 µg of chromatin. |
| ICC/IF | (3) | Use at an assay dependent concentration. |
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
Omim: 602810 Human
SwissProt: P68431 Human
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
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 (mono methyl K9) antibody - ChIP Grade (ab8896)
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab8896 (blue), and 20µl 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 H3 (mono methyl K9) antibody - ChIP Grade (ab8896)This image is courtesy of an anonymous Abreview
Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896) at 1/300 dilution (Incubated for 10 minutes at 20°C. Diluent buffer used was 0.5% milk in TBS, 0.1% Tween.) + HUH-7 cells at 15 µg
Secondary
Sheep anti-rabbit IgG (HRP) at 1/20000 dilution
Predicted band size: 15 kDa
Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896)
All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation
Lane 2 : Calf Thymus Histone Preparation with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg/ml
Lane 8 : Calf Thymus Histone Preparation with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
Lane 9 : Calf Thymus Histone Preparation with Human Histone H3 (unmodified) peptide (ab7228) at 0.5 µg/ml
Lane 10 : Calf Thymus Histone Preparation with Human Histone H3 (unmodified ) peptide (ab2623) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDawhy is the actual band size different from the predicted?
ab8896 specifically recognises mono-methyl K9 (lane 1) in calf thymus lysate at 17kDa. ab8896 is successfully blocked using the immunizing peptide (lane 2 ab1771), but not the di-methyl K9 (lane 3 ab1772), the tri-methyl K9 (lane 4 ab1773), the mono-methyl K27 (lane 5 ab1780), the di-methyl K27 (lane 6 ab1781), the tri-methyl K27 (lane 7 ab1782), the mono-methyl K4 (lane 8 ab1340) nor the corresponding unmodified peptides (lane 9 ab7228, lane 10 ab2623). This implies that ab8896 is specific to mono-methyl K9.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896)This image is courtesy of an anonymous Abreview
Ab8896 staining Histone H3 (mono methyl K9) in HeLa (Cervical cancer cell line) cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton/TBS and blocked with 2% serum for 30 minutes at 25°C. Samples were incubated with primary antibody at 1/100 dilution for 1 hour at 25°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit was used as a secondary antibody at a 1/1000 dilution.
Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896)This image is courtesy of an anonymous Abreview
Diluent: TBS + 0.1% Tween 20 + 5% BSA. Incubated for 12 hours at 4°C + HeLa whole cell lysates at 20 µg
Secondary
Anti-rabbit IgG, HRP-linked Antibody at 1/2000 dilution
Predicted band size: 15 kDa
Exposure time: 1 minute
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896)This image is courtesy of an Abreview submitted by Kirk Mcmanus.
Ab8896 staining Histone H3 (mono methyl K9) in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.05% Triton X100 in PBS. Samples were incubated with primary antibody at 1/200 dilution in PBS for 1 hour at 22°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit was used as the secondary antibody at a 1/200 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896)
ICC/IF image of ab8896 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 ab8896 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was a goat anti-rabbit Alexa Fluor® 488 (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 1.43µM for 1hour at room temperature.
Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896)This image is courtesy of an anonymous abreview.
All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896) at 1/1000 dilution
Lane 1 : Nuclear lysates prepared from Hela cell: SiRNA untreated
Lane 2 : Nuclear lysates prepared from Hela cells treated with G9a SiRNA
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Alkaline Phosphatase conjugated rabbit polyclonal rabbit IgG (Fc) at 1/5000 dilution
Predicted band size: 15 kDa
Observed band size: 16 kDawhy is the actual band size different from the predicted?
Exposure time: 10 minutes
