Swift™ ChIP Kit (MeDIP) (VS-0725-FY4)
CAT#: VS-0725-FY4
The MeDIP (Methylated DNA Immunoprecipitation) ChIP Kit provides a robust solution for enriching methylated DNA directly from cells. Recognizing the critical role of DNA methylation (5-mC) in gene expression, tumorigenesis, and various diseases, this kit utilizes a specific anti-5-mC antibody to immunoprecipitate methylated genomic DNA. The enriched fractions are ideal for diverse downstream applications, including qualitative and quantitative PCR, Southern blot, and DNA microarray analysis. This enables researchers to investigate DNA methylation patterns, offering valuable insights into their association with gene regulation and cancer.
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
- 24 Tests
- Components
- 8-Well Assay Strips (with Frame): 3 x 1 Unit
8-Well Strip Caps: 3 x 1 Unit
Binding Buffer: 1 x 5 mL
ChIP Dilution Buffer: 1 x 4 mL
DNA Release Buffer: 1 x 2 mL
Elution Buffer: 1 x 0.6 mL
F-Collection Tube: 30 x 1 Unit
F-Spin Column: 30 x 1 Unit
Lysis Buffer: 1 x 4 mL
Pre-Lysis Buffer: 1 x 4 mL
Proteinase K (10 mg/mL): 1 x 25 µL
Reverse Buffer: 1 x 2 mL
Wash Buffer: 1 x 28 mL
Reagent composition can vary by lot. Contact us for further parameter details.
- Positive Control
- Anti-5-Methylcytosine: 1 x 25 µL
Antibody Buffer: 1 x 15 mL
- Negative Control
- Normal Mouse IgG: 1 x 10 µL
- Biological Samples
- Suspension cells, Adherent cells
- 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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