The EpiQuik™ Plant ChIP Kit is a convenient package of tools that allows the experimenter to investigate protein-DNA interaction in vivo efficiently. The entire procedure can be completed within 6 hours and produces far superior results than any competitor kits. The EpiQuik™ Plant 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 6 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.
See also a quick chart to compare ChIP kits.
Protein-DNA interaction play a critical role for cellular functions such as signal transduction, gene transcription, chromosome segregation, DNA replication and recombination, and epigenetic silencing. In plants, interactions between the DNA-binding proteins and cognate promoter sequences are primary determinants in establishing spatial and temporal expression patterns of genes that effect homeostasis, development, and adaptation. Chromatin Immunoprecipitation (ChIP) offers an advantageous tool for identifying direct genomewide associations between specific regulatory proteins and their target genes. Unlike other methods such as EMASA, DNA microarrays, and report gene assays, which analyze direct interactions between protein and DNA in vitro, ChIP can detect that a specific protein binds to the specific sequences of a gene in living cells.
Principle & Procedure
This ChIP kit includeds all reagents required for carrying out a successful chromatin immunoprecipitation from plant cells. Particularly, this kit includes a ChIP-grade dimethyl-histone H3-K9 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 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 plant tissue (flowers, leaves, young seedlings). In general, the input amount should be from 20 to 50 mg of plant tissue for each reaction.
CP1 (Wash Buffer)
CP2 (Antibody Buffer)
CP3C (5X Lysis Buffer I)
CP3D (Lysis Buffer II)
CP3E (Lysis Buffer III)
CP3F (Lysis Buffer IV)
CP4 (ChIP Dilution Buffer)
CP5 (DNA Release Buffer)
CP6 (Reverse Buffer)
CP7 (Binding Buffer)
CP8 (Elution Buffer)
Protease Inhibitor Cocktail (100X)*
Normal Mouse IgG (1 mg/ml)*
Anti-Dimethyl H3-K9 (1 mg/ml)*
Proteinase K (10 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.