In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product is observed in the experimental sample, add more DNA to the PCR reaction or increase the number of amplification cycles. Furthermore, if you have any problem with antibodies, make sure to use the ChIP-validated antibody.
Get tips on using Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) to perform ChIP Anti-bodies H3K27me3
Get tips on using Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade to perform ChIP H3K27me3 - Sheep Rat YFP Tag
Get tips on using Bovine Endothelial Cell Media to perform Mammalian cell culture media BAOEC
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Get tips on using Anti-trimethyl-Histone H3 (Lys27) Antibody to perform ChIP Anti-bodies H3K27me3
Get tips on using Anti-trimethyl-Histone H3 (Lys27) Antibody to perform ChIP H3K27me3 - Sheep Rat YFP Tag
Get tips on using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733 to perform ChIP Anti-bodies H3K27me3
When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.
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