High Throughput ChIP-IT® Kit

Synonyms

High Throughput ChIP-IT, 53146, chipit, chip it, chip, ChIP, magnetic ChIP, chromatin IP, chromatin immunoprecipitation, magnetic beads, high throughput, high-throughput

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Overview

High Throughput ChIP-IT^® Kit Overview

The High Throughput ChIP-IT^® Kit provides an optimized protocol and all the reagents needed to perform ChIP assays for histone modifications for transcription factors from 24 cell or tissue samples, yielding qPCR- or next-generation sequencing-ready samples with much less hands-on time than traditional methods.

This kit is designed to work with chromatin sheared with the PIXUL™ Multi-Sample Sonicator to give a robust, streamlined workflow compatible with processing multiple samples as efficiently as possible.

The kit works with chromatin samples prepared by other methods as well. Simply dilute your samples with the 5X ChIP Buffer provided in the kit and they are ready for High Throughput ChIP-IT^® processing.

High Throughput ChIP-IT^® Kit Advantages

• Optimized protocol and reagents ensure efficient results
• Streamlined workflow reduces both total time and hands-on time relative to traditional ChIP protocols
• Low chromatin input required, 70 ng (10,000 cell equivalents) for histone ChIP, 333 ng (50,000 cells) for transcription factor ChIP
• Optimized reverse-crosslinking step in only 2 hours

Contents

High Throughput ChIP-IT^® Kit Contents

The kit contains sufficient reagents for 24 ChIP reactions. The reagents in the kit have multiple storage temperatures. Please store components accordingly. All reagents are guaranteed stable for 6 months from date of receipt when stored properly. Do not re-freeze Protein G Magnetic Beads.

• Protein G Magnetic Beads; Store at 4°C
• TE, pH 8.0; Store at RT
• Protease Inhibitor Cocktail; Store at -20°C
• Blocking Agent AM1; Store at -20°C
• BSA (10 mg/mL); Store at -20°C
• 5X ChIP Buffer; Store at RT
• ChIP Buffer; Store at RT
• Blocker; Store at -20°C
• Wash Buffer AM1; Store at RT
• LiCl Buffer; Store at RT
• Elution Buffer AM4; Store at RT
• Proteinase K (10 µg/µL); Store at -20°C
• RNase A (10 µg/µL); Store at -20°C
• Low EDTA TE; Store at RT
• SPRI Beads; Store at 4°C
• Bar Magnet and Glue Dots; Store at RT

Data

High Throughput ChIP-IT^® Kit Data

Figure 1: Automated chromatin immunoprecipitation (ChIP) was performed on a Beckman Coulter Biomek i7 Automated Workstation using the High Throughput ChIP-IT^® Kit with 30 µg of PIXUL™ Multi-Sample Sonicator-generated chromatin from the MM1S and KMS11 multiple myeloma cell lines immunoprecipitated with 4 µg of AbFlex^® Histone H3K4me3 antibody (Cat. No. 91264). ChIP DNA was sequenced on the Illumina^® NextSeq^® 500 Instrument and sequence tags were mapped to identify Histone H3K4me3 binding sites. Here a region of chromosome 19 (56,000,000:56,600,000) is shown, indicating 3 KMS11-specific H3K4me3 peaks. Top row, MM1S; middle row, KMS11; bottom, pooled input.

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Figure 2: 4.8 μg of K562 chromatin sonicated by the PIXUL™ Multi-Sample Sonicator were used in ChIP reactions with antibodies to histone H3K4me3 and histone H3K27ac using the High Throughput ChIP-IT^® Kit. Purified DNA from the ChIP reactions and inputs were analyzed by qPCR with primers for GAPDH (darker shade portion of bars) and an untranslated, negative control genomic locus (lighter shaded portion of bars).

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