Antibody

id 87
Antibody name H3K27me3
Host organism Rabbit
Poly-/Monoclonal Polyclonal
Target H3K27me3
Peptide TKAARK(me3)SAPATG
Reacts with human, broad range expected
Specificity Testing
H3K27me3, lot A1811-001P ChIP assay
ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K27me…
Chromatin Immunoprecipitation
Recommendation Recommended
Dilution 1 µg/ChIP
Chromatin Amount 100 000 cells
Chromatin Source K562 cells
Capture Method protein A coated magnetic beads
Protocols fixed chromatin
Protocol references diagenode.com/en/catalog/chromatin-function-74/chromatin-immunoprecipitation-75/manual-chip-kits-77/product/…
Comments This antibody was tested on 10e6 HeLa cells using the Diagenode AutoHistone ChIP-seq kit (AB-Auto02-A100) and on 10e5 K562 cells using the Diagenode iDeal ChIP-seq kit (AB-001-0024). 1 µg of antibody was used in both assays.
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
H3K27me3, lot A1811-001P ChIP-seq profile, MYT1 p…
ChIP-seq was performed with the antibody against H3K27me3 and the IP'd DNA was analysed o…
H3K27me3, lot A1811-001P ChIP-seq profile, TSH2B …
ChIP-seq was performed with the antibody against H3K27me3 and the IP'd DNA was analysed o…
H3K27me3, lot A1811-001P ChIP-seq profile, chromo…
ChIP-seq was performed with the antibody against H3K27me3 and the IP'd DNA was analysed o…
ChIP-sequencing
Recommendation Recommended
Dilution 1 µg/ChIP
Chromatin Amount 100 000 cells
Chromatin Source human (K562 cells)
Capture Method protein A coated magnetic beads
Protocols fixed chromatin
Protocol references iDeal ChIP-seq kit (Diagenode cat. No. AB-001-0024; diagenode.com/en/catalog/chromatin-function-74/chromatin-immunoprecipitation-75/manual-chip-kits-77/product/…)
Comments ChIP was performed on sheared chromatin from 100 000 K562 cells using 1 µg of the antibody against H3K27me3 with the "iDeal ChIP-seq" kit (Diagenode, cat. No. AB-001-0024).
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
H3K27me3, lot A1811-001P dot blot analysis
To test the cross reactivity of the Diagenode antibody against H3K27me3, a Dot Blot analy…
Dot blot
Recommendation Recommended
Dilution 1/20000
Blocker TBS-T containing 5% skimmed milk
Incubation 1 hour at room temperature
Secondary Ab ECL peroxidase labelled anti-rabbit antibody (GE Healthcare)
Comments Dot blot was performed for 1 hour at room temperature in TBS-T containing 5% skimmed milk. An ECL peroxidase labelled anti-rabbit antibody (GE Healthcare) was used as secondary antibody
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
H3K27me3, lot A1811-001P, ELISA result
To determine the titer, an ELISA was performed using a serial dilution of the Diagenode a…
Enzyme Linked Immunosorbent Assay
Recommendation Recommended
Dilution dilution curve
Blocker PBS-T containing 3% BSA
Incubation overnight, 4°C
Secondary Ab Goat Anti Rabbit HRP conjugated (Sigma)
Comments ELISA was performed to determine the titer of the antibody.
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
H3K27me3, lot A1811-001P IF assay
NIH3T3 cells were stained with the H3K27me3 antibody(lot A1811-001P) and with DAPI. Cells…
Immunofluorescence
Recommendation Recommended
Dilution 1/500
Fixation 4% formaldehyde in PBS, 10 min RT
Permeabilization 0.5% Triton X-100 in PBS, 15 min RT
Blocker PBS containing 5% normal goat serum, 1% BSA, 0.1% Triton X-100
Incubation 4°C overnight
Secondary Ab Goat Anti-Rabbit Alexa 488, Invitrogen (A11008)
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
H3K27me3, lot A1811-001P WB assay
Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lan…
Western blot
Recommendation Recommended
Dilution 1/1000
Blocker TBS-T containing 5% skimmed milk
Incubation overnight, 4°C
Secondary Ab ECL peroxidase labelled anti-rabbit antibody (GE Healthcare)
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
H3K27me3, lot A1811-001P peptide array
The specificity of the antibody was further demonstrated by peptide array analyses on an …
H3K27me3, lot A1811-001P, ranked list of peptides
Normalized intensities of the different peptides present on the array.
Peptide array
Recommendation Recommended
Comments Peptide array was performed for 1 hour at room temperature in TBS-T containing 5% skimmed milk. The antibody was used at a dilution of 1/20000.
Link to antibody source diagenode.com/en/catalog/antibodies-3/histone-h3-and-modified-h3-27/product/…
Supplier
Company: Diagenode
Lot no: A1811-001P
Catalog no: pAb-195-050
Website: http://www.diagenode.com
Contact email: info@diagenode.com
This data is online since 2012-07-02 14:47
Last update 2013-04-25 12:37
provided by Jan Hendrickx