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.
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-trimethyl-Histone H3 (Lys4) Antibody, clone MC315, rabbit monoclonal to perform ChIP Anti-bodies H3K4me3
Get tips on using CTCF (D31H2) XP® Rabbit mAb #3418 to perform ChIP Anti-bodies CTCF
Get tips on using Estrogen Receptor α (D8H8) Rabbit mAb #8644 to perform ChIP Anti-bodies ERα
Get tips on using HDAC1 (D5C6U) XP® Rabbit mAb #34589 to perform ChIP Anti-bodies HDAC1
Get tips on using Bcl-11B (D6F1) XP® Rabbit mAb #12120 to perform ChIP Anti-bodies CtIP/BCL11A
Get tips on using Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684) to perform ChIP Anti-bodies H3K27me2
Get tips on using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 to perform ChIP Anti-bodies H3K9-Ac
Get tips on using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb #9725 to perform ChIP Anti-bodies H3K4me2
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