MAGnify™ Chromatin Immunoprecipitation System
MAGnify™ Chromatin Immunoprecipitation System
Applied Biosystems™

MAGnify™ Chromatin Immunoprecipitation System

The MAGnify™ Chromatin Immunoprecipitation System provides a streamlined, optimized assay for the enrichment of chromatin/protein complexes and DNA recovery usingRead more
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Catalog number 492024
Price (USD)
834.00
Each
Add to cart
Price (USD)
834.00
Each
Add to cart
The MAGnify™ Chromatin Immunoprecipitation System provides a streamlined, optimized assay for the enrichment of chromatin/protein complexes and DNA recovery using magnetic bead capture technology. The isolated DNA is ready for downstream analysis by methods such as PCR- or qPCR-based assays, or massive parallel DNA sequencing.

ChIP
Chromatin immunoprecipitation (ChIP) is a powerful technique for studying the association of certain proteins with specific regions of the genome. These sequence-specific DNA-binding proteins are believed to play a role in such cellular processes as DNA replication, recombination, repair, and segregation; chromosomal stability; cell-cycle progression; and epigenetic silencing. In a standard ChIP assay, a cell is fixed via formaldehyde treatment and the chromatin is sheared and immunoprecipitated via a highly specific antibody. The researcher then analyzes the DNA to identify the genomic regions where the chromatin-associated proteins bind to the chromatin in vivo. This kit enables researchers to start with lower sample amounts than traditional ChIP workflows, thereby preserving precious samples, and the protocol can be completed in a single day, compared with 2–3 days for a traditional ChIP assay. The kit can be used with our suite of ChIP-validated antibodies, and is also complementary with MethylCode™ and NCode™ products for downstream epigenetics research.

Using the MAGnify™ system
When using the MAGnify™ system, you treat cells or tissue with formaldehyde to generate protein-protein and protein-DNA crosslinks between molecules in close proximity within the chromatin complex. The cells are then lysed, and the chromatin is released from the nuclei and sheared by sonication to reduce the average DNA fragment size to 200–500 bp for analysis by quantitative real-time PCR (qPCR) or 100–300 bp for analysis by massive parallel DNA sequencing. You then immunoprecipitate and isolate the crosslinked protein of interest using a specific ChIP-qualified antibody conjugated to Dynabeads™ Protein A/G. The formaldehyde crosslinking is reversed by heat treatment, and the DNA associated with that protein is purified. The DNA is now ready for downstream analyses such as end-point PCR or quantitative PCR (qPCR), genome-wide analyses using promoter-tiling arrays, or next-generation sequencing. In PCR/qPCR analysis, primers are designed to span the desired DNA sequence of interest, and the data demonstrates whether the specific protein of interest is associated in vivo with that DNA region.
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For Research Use Only. Not for use in diagnostic procedures.
Specifications
High-throughput CompatibilityHigh-throughput Compatible
Sample TypeCell Cultures, DNA (Genomic), Live Cells
Product LineMAGnify™
Quantity1 Kit
For Use With (Application)Chromatin Immunoprecipitation
TypeImmunoprecipitation/Co-Immunoprecipitation Kit
Sufficient For24 Reactions
Unit SizeEach
Contents & Storage
Module 1 (shipped on wet ice, store at 4°C):
• 2 × 1 ml Glycine (1.25 M)
• 250 μl Dynabeads™ Protein A/G (do not freeze)
• 1.4 ml Reverse Crosslinking Buffer
• 500 μl DNA Purification Magnetic Beads (do not freeze)
• 1.4 ml DNA Purification Buffer
• 200 μl Proteinase K (20 mg/ml)

Module 2 (shipped on wet ice, store at 4°C):
• 10 ml IP Buffer 1
• 7.5 ml IP Buffer 2
• 8 ml DNA Wash Buffer
• 7.2 ml DNA Elution Buffer

Module 3 (shipped on dry ice, store at -20°C):
• 100 μl Protease Inhibitors (200X)
• 15 μl Mouse IgG (1 μg/μl)
• 15 μl Rabbit IgG (1 μg/μl)

Module (shipped on dry ice, store at -20°C):
• 8 ml Dilution Buffer
• 3.6 ml Lysis Buffer

Frequently asked questions (FAQs)

What is the recommended amount of starting material when working with the MAGnify Chromatin Immunoprecipitation System kit?

For each ChIP reaction, we recommend using 10,000-300,000 cells or 0.167-5 mg of tissue. To ensure consistency and decrease experimental variability, we recommend preparing a common chromatin batch suitable for multiple ChIP experiments. Note that following lysis, samples are at a concentration of 1 million cells/50 µL.

I am getting equivalent PCR signal from positive and negative control IP samples. Why is this?

There might be excess chromatin or antibody added to the IP, or insufficient amount of DNA template added into the PCR reaction. Also your PCR conditions might need optimizing. Try decreasing the number of amplification cycles in PCR. Finally, it is ideal to have duplicate or triplicate runs for each IP to identify any issues, like human or product errors.

I am not getting PCR signal in the experimental IP samples, while the results from positive and negative control IPs are as expected. What happened?

The most possible cause is the antibody does not function in IP. Not all antibodies used for western blotting will work well in ChIP. You need to verify the antibody is qualified for ChIP or IP applications. And try adding more antibody to the IP reaction and more DNA template to the PCR.

I am not getting any PCR signal with the positive control IP samples. Do you have some tips?

This is likely to be caused by insufficient chromatin amount in the IP reaction or insufficient antibody incubation time. We also recommend optimizing the crosslinking condition, and monitoring sonication of nuclei by microscope to ensure complete lysis.

I am doing chromatin analysis and am not getting PCR signal in the total input control samples. What should I do?

First make sure that the PCR condition is fully optimized and check the primer design. Then try increasing the amount of template DNA added to the PCR reaction. Finally, evaluate sonication of nuclei by microscope to ensure complete lysis.