Antibody

id 101
Antibody name H3K27ac
Host organism Rabbit
Poly-/Monoclonal Polyclonal
Target H3K27ac
Peptide TKAARK(ac)SAPATG
Reacts with human, broad range expected
Specificity Testing
H3K27ac, lot A1723-0041D ChIP assay, 10e6 HeLa ce…
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K27ac…
H3K27ac, lot A1723-0041D ChIP assay, 10e5 K562 ce…
ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K27ac…
Chromatin Immunoprecipitation
Recommendation Recommended
Dilution 1-2 µg/ChIP
Chromatin Amount 1 000 000 cells/100 000 cells
Chromatin Source human cell lines (HeLa, K562)
Capture Method protein A coated magnetic beads
Protocols fixed chromatin
Protocol references Auto Histone ChIP-seq kit (Diagenode cat. No. AB-Auto02-0100; diagenode.com/en/catalog/chromatin-function-74/chromatin-immunoprecipitation-75/automated-chip-kits-76/product/…); iDeal ChIP-seq kit (Diagenode cat. No. AB-001-0024)
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).
Link to antibody source diagenode.com/en/catalog/antibodies-3/chip-seq-grade-antibodies-97/product/…
H3K27ac, lot A1723-0041D, ChIP-seq profile along …
ChIP was performed on sheared chromatin from 100,000 K562 cells using 1 µg of the Diageno…
H3K27ac, lot A1723-0041D ChIP-seq profile, EIF4A2…
ChIP was performed on sheared chromatin from 100,000 K562 cells using 1 µg of the Diageno…
H3K27ac, lot A1723-0041D ChIP-seq profile, GAPDH …
ChIP was performed on sheared chromatin from 100,000 K562 cells using 1 µg of the Diageno…
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/…)
Link to antibody source diagenode.com/en/catalog/antibodies-3/chip-seq-grade-antibodies-97/product/…
H3K27ac, lot A1723-0041D, dot blot analysis
To test the cross reactivity of the Diagenode antibody against H3K27ac, a Dot Blot analys…
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/chip-seq-grade-antibodies-97/product/…
H3K27ac, lot A1723-0041D 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/chip-seq-grade-antibodies-97/product/…
H3K27ac, lot A1723-0041D IF assay
HeLa cells were stained with the H3K27ac antibody(lot A1723-0041D) and with DAPI. Cells w…
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/chip-seq-grade-antibodies-97/product/…
H3K27ac, lot A1723-0041D Western blot analysis
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/chip-seq-grade-antibodies-97/product/…
Supplier
Company: Diagenode
Lot no: A1723-0041D
Catalog no: pAb-196-050
Website: http://www.diagenode.com
Contact email: info@diagenode.com
This data is online since 2012-09-07 11:39
Last update 2012-10-16 14:34
provided by Jan Hendrickx