		Transcriptional Regulation Co-Immunoprecipitation DNA Repair Luciferase Reporter Assay Nuclear Receptor ELISAs Supershift & Gelshift Assays Transcription Factor ELISA Epigenetics & Chromatin Chromatin Assembly Chromatin IP DNA Damage – Phospho-H2AX DNA Methylation Histone Acetyltransferase Activity (HAT) Histone Deacetylase Activity (HDAC) Histone Modification ELISAs Histone Purification Methylated Histones Protein Modification GTPase Activation Assay Kinase Assay Substrate Mitosis Assays Nitric Oxide Quantitation Phospho-Protein Detection Sandwich ELISAs SUMOylation Antibodies, Extracts & Proteins Antibodies Cell Extracts Recombinant Proteins Fluorescence & Labeling Albumin Quantification Cell Viability Assay Fluorescent Dyes & Labeling Immunofluorescence Live Cell Membrane Staining Mitosis Determination Protein and Extract Quantification Protein Labeling & Translocation Sample Preparation Fluorescent Dye Labeling Mitochondrial Separation mRNA Isolation Normalization Nuclear & Cytoplasmic Extraction Protein & Extract Quantification Transfection & Expression Competent E. coli Gene Silencing Protein Transfection Staining Reagents Bioinformatics & Text Mining TimeLogic® Biocomputing Solutions AKS 2

Universal Magnetic Co-IP Kit

magnetic beads for low background co-immunoprecipitation

The Universal Magnetic Co-IP Kit improves co-immunoprecipitation (Co-IP) through the use of protein G-coated magnetic beads, which speed and simplify the procedure while greatly reducing background. The kit includes optimized reagents for making both nuclear & whole-cell extracts from cells or tissue, giving you the flexibility to Co-IP any protein complex, whether it was originally bound to DNA or in the cytoplasm.

Universal Magnetic Co-IP Kit advantages

Perform Co-IP of nuclear or cytoplasmic protein complexes, beginning from cells or tissue
Magnetic beads speed and simplify the procedure while reducing background, enabling the use of low-stringency buffers that maintain complex integrity
Nuclear extraction procedure removes nuclear protein complexes from the DNA without disrupting protein/protein interactions
Inhibitor solutions included to preserve the integrity of protein modifications

Perform Co-IP of cytoplasmic & nuclear protein complexes

While Co-IP is often used to study cytoplasmic protein complexes, traditional methods are not optimal for studying DNA-binding proteins because these nuclear complexes are fragile and can be difficult to remove from the DNA. This causes them to either be disrupted during extraction, or to be lost because they remain bound to the DNA. For this reason, in addition to containing components for whole-cell extraction, the Universal Magnetic Co-IP Kit provides nuclear extraction reagents that have been optimized to preserve nuclear protein complexes. The reduced background provided by the low non-specific binding of the kit's magnetic beads makes it possible to use a low-stringency Co-IP/Wash Buffer that helps maintain the protein complexes (Figure 1). To further improve Co-IP of nuclear complexes, the kit’s proprietary Enzymatic Shearing Cocktail uses DNA digestion to release undissociated protein complexes from the DNA. Even histone complexes, which are not released from the DNA by traditional high-salt extraction methods, can be studied (Figure 2).

Figure 1: Nuclear Co-IP of SRC-1 and ERα.

The Universal Magnetic Co-IP Kit was used to make nuclear extract from MCF-7 cells induced 1 hour with 10 nM Estradiol. IP was performed on 300 µg samples using 2 µg of SRC-1 pAb, ERα pAb and rabbit IgG (as a negative control). Western blot was then performed using the ERα pAb on 10 µg Input Extract, SRC-1 IP, ERα IP and the rabbit IgG IP.

Figure 2: Detection of acetylated Histone H3.

HeLa nuclear extracts were made using the Universal Magnetic Co-IP Kit and a traditional high-salt extraction protocol, supplemented with 1 µM trichostatin A, a deacetylase inhibitor. Five and ten µg samples of each extract were used in Western with Histone H3 acetyl pAb. Protein was detected only in samples made using the kit’s nuclear extraction procedure, as it was designed to release histone and other protein complexes from DNA while preserving modifications.

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