Swift™ Plus Sonication ChIP Kit (VS-0725-FY12)
CAT#: VS-0725-FY12
The Plus Sonication Chromatin IP Kit is a comprehensive solution for ChIP assays, containing all essential buffers and reagents for up to 24 immunoprecipitations from cells or tissue. Optimized for 4 x 10⁶ cells or 25 mg tissue per IP, the entire assay can be completed in just two days and is suitable for both ChIP-qPCR and ChIP-seq.
The protocol involves formaldehyde fixation, lysis, and chromatin fragmentation via sonication to yield 200-1000 bp fragments. Optimized lysis buffers ensure maximum enrichment of histones, transcription factors, and cofactors. Immunoprecipitations are conducted with ChIP-validated antibodies and ChIP-Grade Protein G Magnetic Beads. Following cross-link reversal, DNA is purified using spin columns. The kit includes crucial controls like Histone H3 Antibody, Normal Rabbit IgG Antibody, and RPL30 primer sets, enabling detection of protein-DNA interactions and histone modifications in mammalian samples.
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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