PMA-treated THP1 cells were infected with 6xHis-Rv2966c::M. bovis BCG at an MOI of 20:1for 24 h, followed by 24 h of rest. Following infection, cells were washed with PBS and cross-linked with 2.7% formaldehyde for 10 min at room temperature. The reaction was quenched with 125 mM glycine for 5 min. Cells were harvested and resuspended in 0.1% SDS. Chromatin was prepared by sonicating the cells in Bioruptor (Diagenode) to obtain DNA fragments in the range of 100–300 bp. For each ChIP, chromatin from 10 million of infected or uninfected cells was used. Chromatin was incubated with 2 μg of anti-His antibody (abcam) or IgG (Rabbit, Diagenode) for 12 h, followed by incubation with Protein A beads for 4 h. All the steps were done using auto ChIP kit (Diagenode) in the IPStar Compact machine (Diagenode). Ten percent chromatin was used as input.
Auto chip kit
Manufactured by Diagenode
The Auto ChIP kit is a laboratory equipment product designed for chromatin immunoprecipitation (ChIP) experiments. The core function of the kit is to automate the ChIP process, providing a streamlined and reproducible workflow for researchers studying DNA-protein interactions.
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Lab products found in correlation
3 protocols using auto chip kit
1
ChIP-seq of Rv2966c in M. bovis BCG-infected THP1 cells
2
ChIP Assay with Automated System
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ChIP was performed using auto ChIP kit (Diagenode) according to manufacturer’s instructions. Anti-human H3K27me3 (pAb-069-050, Diagenode) produced in rabbit and also non-immune rabbit IgG (negative control) (kch-504-250, Diagenode) were used. The ChIP was carried out by SX-8X® Automated System (Diagenode) according to the protocol provided by the manufacturer. The reaction was incubated for 2 h for antibody coating with protein A-coated magnetic beads, then for 10 h at 4°C for immunoprecipitation reaction. Later on, 1 μL of proteinase K was added and the reverse cross-linking was performed for 45 min at 65°C. The immunoprecipitated DNA and input samples were analyzed by real-time qPCR.
3
Chromatin Immunoprecipitation in Drosophila
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Chromatin immunoprecipitation was performed as described previously43 (link). Briefly, 3rd instar larvae was homogenised and then sonicated in Biorupter (Diagenode) Biorupter so that DNA was sheared between 200–500 bp. Chromatin immunoprecipitation was performed on the sonicated sample using auto ChIP kit (Diagenode) in the IPStar automated ChIP machine (Diagenode) using specific antibodies.
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