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 in the experimental, add more DNA to the PCR reaction or increase the number of amplification cycles. Choose an alternate, ChIP-validated antibody if the antibody does not work.
Get tips on using SOLiD™ ChIP-Seq Kit, with ChIP magnet to perform ChIP Human - SH-SY5Y
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 (di methyl K4) antibody - ChIP Grade (ab7766) to perform ChIP Anti-bodies H3K4me2
Get tips on using Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895 to perform ChIP Anti-bodies H3K4me1
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 in the experimental, add more DNA to the PCR reaction or increase the number of amplification cycles. Choose an alternate, ChIP-validated antibody if the antibody does not work.
Get tips on using EZ-ChIP™ to perform ChIP Human - HUVEC
Get tips on using OneDay ChIP kit to perform ChIP Human - Kupffer Cells
Get tips on using ChIP Kit (ab500) to perform ChIP Human - HUH-7
Get tips on using EZ-ChIP™ to perform ChIP Human - HUH-7
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