Anti-Histone H3 (mono methyl K4)抗体- ChIP Grade
参阅全部 Histone H3 一抗
兔多克隆抗体to Histone H3 (mono methyl K4) - ChIP Grade
Rabbit
Specific for mono-methylated Lysine 4 of histone H3. Does not recognise di- or tri-methyl Lysine 4 nor methylation at Lysine 9.
适用于: ICC/IF, ChIP, WB, IHC-Pmore details
与反应: Mouse, Rat, Cow, Human
预测可用于: Pig, Saccharomyces cerevisiae, Tetrahymena, Xenopus laevis, Drosophila melanogaster, Plants, Mammals, Plasmodium falciparum, Xenopus tropicalis, Candida albicans
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab1340)
ChIP: Chromatin prepared from U-2 OS cells; Metastatic clear cell renal carcinoma cells (M1A). WB: Calf thymus histone; HeLa; NIH3T3; PC12 ICC/IF: HeLa cells. IHC-P: Human colon tissue.
Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.
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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: 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
50 µg 浓度为 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab8895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 | (6) | Use a concentration of 1 - 5 µg/ml. Works better if cells fixed in methanol. |
| ChIP | (16) | Use 2 µg for 25 µg of chromatin. We recommend Myo-D ChIP primer pair ab269261 as positive control. |
| WB | (22) | 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). |
| IHC-P | (4) | Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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: 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
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ChIP - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)
Chromatin was prepared from U-2 OS 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 ab8895 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
Western blot - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)
All lanes : Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) at 1 µg/ml
Lane 1 : Calf Thymus Histone at 0.5 µg
Lane 2 : HeLa Nuclear – Triton Prep at 10 µg
Lane 3 : NIH3T3 Nuclear – Triton Prep at 10 µg
Lane 4 : PC12 Nuclear at 10 µg
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
Gel type: MES
Western blot - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)
All lanes : Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) at 1/500 dilution
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml
Lane 3 : Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/ml
Lane 4 : Calf thymus histone lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 1 µg/ml
Lane 5 : Calf thymus histone lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/ml
Lane 6 : Calf thymus histone lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 1 µg/ml
Lane 7 : Calf thymus histone lysate with Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 18 kDawhy is the actual band size different from the predicted?
Exposure time: 2 minutes
ab8895 is specific for mono-methylated Lysine 4 of histone H3 and does not recognize di- or tri-methyl Lysine 4 nor methylation at Lysine 9. This is shown in lane 2 where the activity of the antibody is specifically blocked by the addition of the immunizing peptide (ab1340).
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)
ab8895 staining Histone H3 (mono methyl K4) 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 ab8895 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)
IHC image of ab8895 staining Histone H3 (mono methyl K4) 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 (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8895, 0.5µg/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. 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.
ChIP - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)This image is courtesy of an Abreview submitted by Paulo Rodrigues
ChIP analysis using ab8895 binding Histone H3 (mono methyl K4) in Metastatic clear cell renal carcinoma cells (M1A). Cells were cross-linked for 10 minutes with 1% formaldehyde. Samples were incubated at 1/100 dilution with primary antibody for 12 hours at 4°C.
Positive control:Metastatic clear cell renal carcinoma cells (M1A).
Negative Control:Parental clear cell renal carcinoma cells (786-O).
