Swift™ Enzymatic ChIP Kit (Magnetic Beads) (VS-0725-FY11)
CAT#: VS-0725-FY11
The Enzymatic Chromatin IP Kit (Magnetic Beads) offers a complete solution for chromatin immunoprecipitation assays, including buffers and reagents for up to 6 chromatin preparations and 30 immunoprecipitations. Optimized for 4 x 10⁶ cells per experiment, this kit allows for a full assay completion in just two days. It provides flexibility for scaling experiments up or down based on cell numbers. The process involves fixing cells with formaldehyde, lysing, and fragmenting chromatin using Micrococcal Nuclease to yield 1 to 5 nucleosome fragments. Following protein-DNA cross-link reversal, DNA is purified via spin columns, ensuring high-quality results.
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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