siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line)

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized PC-3

Get tips on using RNAzol® RT to perform RNA isolation / purification Cells - immortalized PC-3

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized PC-3

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized PC-12

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - immortalized PC-3

Get tips on using Direct-zol RNA Kits to perform RNA isolation / purification Cells - immortalized PC-3

Get tips on using Pierce™ BCA Protein Assay Kit to perform Protein quantification Mammalian cells - PC-12

The DNA concentration after using this DNA isolation kit is sometimes too low and thus it is not sufficient for my follow-up experiments. How can I improve it?

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.

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.