Swift™ Enzymatic ChIP Kit (Agarose Beads) (VS-0725-FY14)
CAT#: VS-0725-FY14
The Enzymatic Chromatin IP Kit (Agarose Beads) offers a streamlined solution for chromatin immunoprecipitation, providing all essential buffers and reagents for both chromatin preparation and IP. Designed for use with 4x10⁶ cells per experiment, this comprehensive kit allows for complete assay execution within two days. A key feature is the use of Micrococcal Nuclease for chromatin fragmentation, a method gentler than sonication, which helps maintain superior chromatin quality and preserve antibody epitopes. The kit also includes DNA purification spin columns for efficient recovery of DNA following immunoprecipitation. To ensure successful experiments, control antibodies such as Histone H3 and Normal Rabbit IgG are provided, alongside human and mouse RPL30 gene primer sets.
Specifications
- Application
- ChIP
- Workflow
- 1. Cell/Tissue Preparation & Cross-linking: Cells or tissues are collected, and DNA-protein interactions are fixed using a cross-linking agent like formaldehyde, followed by quenching with glycine.
2. Cell Lysis & Chromatin Shearing: Cells undergo lysis, and chromatin is mechanically sheared (e.g., via sonication) into smaller fragments, typically 100-1000 bp. Sheared chromatin is then separated from debris by centrifugation.
3. Magnetic Immunoprecipitation: Protein A-coated magnetic beads are washed and then incubated with sheared chromatin and a specific antibody (or control IgG) to capture antibody-protein-DNA complexes.
4. Washing: Non-specifically bound components are removed by extensively washing the immunocomplex-bound beads with various buffers.
5. Elution, Decross-linking & DNA Purification: Immunoprecipitated protein-DNA complexes are eluted. Cross-links are reversed by high-temperature incubation with proteinase K, releasing the DNA, which is then purified.
6. Downstream Analysis: The purified DNA is analyzed to identify and quantify specific bound sequences using methods such as quantitative PCR (qPCR) or Next-Generation Sequencing (ChIP-seq).
- Size
- 1 kit
- Components
- Lysis Buffers
Wash Buffers
Crosslinking Reagents (e.g., formaldehyde; often not directly supplied in kits but required per protocol)
Chromatin Shearing Reagents / Enzymes (e.g., Micrococcal Nuclease or components for sonication)
Proteinase K
DNA Purification Columns or Magnetic Beads
For comprehensive parameter information concerning our buffers, please reach out to us.
- Positive Control
- Rabbit anti-Histone H3 antibody
- Negative Control
- Normal Rabbit IgG
- Primer
- Human RPL30 Exon 3 Primers: 1 x 150 µl
Mouse RPL30 Intron 2 Primers: 1 x 150 µl
- Shipping Conditions
- Blue Ice
- Storage
- All components within this kit remain stable for at least 6 months beyond the reference date on their labels, provided they are stored at the recommended temperature and remain unused.
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