Free of charge vial of the New SimpleChIP Universal qPCR Master Mix
Sonication or Enzymatic Digestion: We have kits to support both experimental methods
Sonication is a popular choice for preparing chromatin for ChIP. However, most protocols require subjecting the chromatin to harsh, denaturing conditions (i.e., high heat, detergent, and shearing force) that can damage both antibody epitopes and the integrity of the chromatin.
CST provides multiple options for avoiding the problems of traditional sonication protocols:
- Sonication: Our sonication protocol uses specially formulated cell and nuclear lysis buffers. This approach protects chromatin integrity and antibody epitopes, resulting in increased ChIP efficiency and making it more compatible for use with transcription factors and cofactors, which are labile and have less stable DNA interactions.
- Enzymatic digestion: Enzymatic digestion is an alternative method of chromatin fragmentation that is compatible with all target types and highly amenable to the immunoprecipitation of transcription factors and cofactors. This approach uses micrococcal nuclease to cut the linker region between nucleosomes. Additionally, it is simple to control and protects chromatin and antibody epitopes from shearing or denaturation, making it an ideal option for users who are new to performing ChIP.
*** The SimpleChIP® Universal qPCR Master Mix is an optimized 2X reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR/FAM channel of most real-time qPCR instruments. It contains Hot Start Taq DNA Polymerase and has been formulated with a unique passive reference dye that is compatible across a variety of instrument platforms (including those that require a high or low ROX reference signal). It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection.
This product is provided in 1 ml volumes sufficient for preparation of 100 qPCR reactions, and is compatible with both enzymatic and sonication-fragmented DNA samples from SimpleChIP®enzymatic and sonication ChIP kits. This master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification and quantitation of DNA, except primers and a DNA template.
ChIP-seq Validated Antibodies
Choosing an antibody that recognizes the correct target protein and does not show non-specific binding with other DNA-binding proteins is critical for a successful ChIP-seq experiment. CST now tests and validates recombinant rabbit monoclonal antibodies in ChIP-seq, because good performance in ChIP-qPCR does not necessarily mean an antibody will perform well for ChIP-seq, which requires more extensive capture of the target protein across a large number of gene loci. CST’s ChIP-seq approved antibodies are tested using the SimpleChIP Enzymatic Chromatin IP protocol followed by next-generation sequencing (NGS).
Advantages of using ChIP-seq antibodies produced at CST are:
- All ChIP-seq approved antibodies from CST are recombinant rabbit monoclonal antibodies, providing greater lot-to-lot reproducibility
- Antibodies are validated and optimized using the SimpleChIP Enzymatic Chromatin IP protocol, saving you time and money
- Antibodies are tested and validated for target specificity across multiple applications, providing reduced non-specific binding and high signal-to-noise ratio in ChIP-seq
- Antibodies are compatible with other ChIP-seq protocols, allowing for use in ChIP kits from CST or other suppliers, and in customized lab protocols
- Expert technical support is available from the same scientists who developed and validated the ChIP-seq antibodies