The EpiQuik™ Tissue Acetyl-Histone H3 ChIP Kit is a convenient package of tools that allows the experimenter to investigate interactions of histone H3 acetylation and DNA in variety of mammalian tissues. The entire procedure can be completed within 5 hours and produces far superior results than any competitor kits. The EpiQuik™ Tissue Acetyl-Histone H3 ChIP kit is suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, ChIP-Seq, and ChIP-on-chip. This kit has the following advantages:
- The fastest procedure available, which can be finished within 5 hours.
- Strip microwell format makes the assay flexible: manual or high throughput.
- Columns for DNA purification are included: save time and reduce labor.
- Compatible with all DNA amplification-based approaches.
- Simple, reliable, and consistent assay conditions.
Histone acetylation takes place at the α-amino groups of conserved lysine residues, where histone acetyltransferases (HATs) are responsible for catalyzing this modification. It has been demonstrated that histone H3 acetylation serves as an epigenetic marker of chromosomal domains. Histone H3 acetylation regulates various cellular physiological processes, including transcriptional activation of genes, chromatin assembly, cell proliferation, and some pathological processes such as tumorigenesis. Chromatin Immunoprecipitation (ChIP) is a powerful technique for studying protein-DNA interaction in vivo. ChIP also offers an advantageous tool that allows identification of activated genes associated with acetylated histone H3. ChIP coupled with microarrays could be further used for profiling or mapping histone H3 acetylation patterns.
Principle & Procedure
This ChIP kit contains all reagents required for carrying out a successful chromatin immunoprecipitation for acetyl-histone H3 from mammalian cells. Particularly, this kit includes a ChIP-grade acetyl-histone H3 antibody and a negative control normal mouse IgG. Chromatin from the cells is extracted, sheared and added into the microwell immobilized with the antibody. DNA is released from the antibody-captured acetyl-histone H3 protein-DNA complex, reversed and purified through the specifically designed F-Spin Column. Eluted DNA can be used for various down-stream applications.
Starting materials can include various tissue samples. In general, the input amount should be 20 mg of tissue for each reaction.
CP1 (Wash Buffer)
CP2 (Antibody Buffer)
CP3 (Lysis Buffer)
CP4 (ChIP Dilution Buffer)
CP5 (DNA Release Buffer)
CP6 (Reverse Buffer)
CP7 (Binding Buffer)
CP8 (Elution Buffer)
Protease Inhibitor cocktail (100X)*
Proteinase K (10 mg/ml)*
Normal Mouse IgG (0.5 mg/ml)*
Anti-Acetyl-Histone H3 (1 mg/ml)*
8-Well Assay Strips (with Frame)
8-Well Strip Caps
* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.