ChIP H3K36Me3 Canine Rabbit

Get tips on using EpiTect ChIP OneDay Kit to perform ChIP Human - HUH-7

Get tips on using Androgen Receptor (D6F11) XP® Rabbit mAb #5153 to perform Immunohistochemistry Human - AR

Get tips on using Lab Vision™ Ki-67, Rabbit Monoclonal Antibody to perform Immunohistochemistry Mouse - Ki67

Get tips on using EpiTect ChIP OneDay Kit to perform ChIP Human - MIA PaCa-2

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.

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

Get tips on using Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660 to perform Immunohistochemistry Human - GR/glucocorticoid receptor

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - MCF7

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - A172

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - A375