ChIP acH4 Mouse Rabbit

Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Human - MIA PaCa-2

Get tips on using Pierce™ Magnetic ChIP Kit to perform ChIP Human - Fibroblast cell lines

Get tips on using Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb #2078 to perform Western blotting IKK Alpha

Get tips on using Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb #4806 to perform Western blotting NF-kB

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 Anti-PI3-kinase p85-α antibody produced in rabbit to perform Autophagy assay cell type - HepG2

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

Get tips on using Phospho-eIF2α (Ser51) (D9G8) XP® Rabbit mAb #3398 to perform Autophagy assay cell type - HEK 293

Get tips on using Recombinant Anti-beta Catenin antibody [E247] - ChIP Grade (ab32572) to perform Immunohistochemistry Mouse - β-Catenin

Get tips on using ChIP-IT® Express Shearing Kits to perform ChIP Human - SW480