Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)
Key features and details
- Rabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade
- Suitable for: PepArr, Dot blot, ChIP, ChIP-sequencing, WB, ICC/IF
- Reacts with: Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade
See all Histone H3 primary antibodies -
Description
Rabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: PepArr, Dot blot, ChIP, ChIP-sequencing, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human, Recombinant fragment
Predicted to work with: Mouse, a wide range of other species
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Immunogen
Synthetic peptide corresponding to Human Histone H3 (tri methyl K9) conjugated to keyhole limpet haemocyanin.
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Positive control
- HeLa whole cell lysate; HeLa histone extract; HeLa cells; Chromatin prepared from HeLa and K562 cells.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservatives: 0.05% Sodium azide, 0.05% Proclin 300
Constituent: 99% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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ChIP-sequencing - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)
ChIP-seq results obtained with ab195412 directed against Histone H3 (tri methyl K9).
ChIP was performed on sheared chromatin from 100,000 K562 cells using 1 μg of ab195412. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the BWA algorithm. The top panel shows the Histone H3 (tri methyl K9) signal distribution along the long arm of human chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The bottom panel shows the signal distribution in a 200 kb region from chromosome 12 surrounding the ZNF12 gene.
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ChIP - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)ChIP results obtained with ab195412 directed against Histone H3 (tri methyl K9).
ChIP assays were performed using human K562 cells, ab195412 and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration consisting of 0.2, 0.5, 1 and 2 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH, c-fos and EIF4A2, used as negative controls, and for ZNF510 and the Sat2 satellite repeat, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)Immunofluorescent analysis of HeLa cells labeling Histone H3 (tri methyl K9) using ab195412 at 1/500 dilution. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the ab19541 (top) diluted 1/500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown at the bottom.
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Peptide Array - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)The specificity of ab195412 was demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. ab195412 was used at a dilution of 1/2,000. Figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.
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Dot Blot - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)Dot Blot analysis was performed to test the cross reactivity of ab195412 against Histone H3 (tri methyl K9) with peptides containing other histone modifications and the unmodified H3K9. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1/20,000. Figure shows a high specificity of the antibody for the modification of interest.
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Western blot - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412)All lanes : Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab195412) at 1/1000 dilution
Lane 1 : HeLa whole cell extract at 25 µg
Lane 2 : HeLa histone extract at 15 µg
Lane 3 : recombinant histone H2A at 1 µg
Lane 4 : recombinant histone H2B at 1 µg
Lane 5 : recombinant histone H3 at 1 µg
Lane 6 : recombinant histone H4 at 1 µg
Predicted band size: 15 kDaantibody diluted in TBS-Tween containing 5% skimmed milk
