CNMCS Compartmental Protein Extraction Kit

Overview

BioChain’s CNMCs Compartmental Protein Extraction Kit is designed for rapid and efficient sequential isolation of cytoplasmic, nuclear, membrane, and cytoskeletal proteins, all from the same tissue or cell samples. The protein extracts isolated using this kit are pure and overcome a major challenge of complex protein separation for downstream functional proteomic analysis.

Description

One of the many challenges in the field of proteomics is separation of complex protein mixtures for quantitative and differential localization analysis. To directly address this issue, BioChain is pleased to offer the CNMCs Compartmental Protein Extraction Kit. This kit is designed for rapid and efficient sequential isolation of cytoplasmic, nuclear, membrane, and cytoskeletal proteins, all from the same tissue or cell samples.

 

The system utilizes our proprietary buffers that have been optimized to prepare the highest quality compartmental protein extractions. The kit also includes our proprietary protease inhibitor cocktail to prevent protein degradation. In conjunction, these components result in protein extracts that may be used for downstream applications such as SDS-PAGE, isoelectric focusing and 2-D gel electrophoresis, Western blotting, gel mobility shift analysis, and other protein assays.

 

Isolating proteins from different subcellular compartments is essential for proteomics research. Many proteins of interest in biomarker discovery are abundant in low quantities. Therefore, their enrichment through fractionation is important to facilitate their study. With our CNMCs kit, scientists will be more readily able to research transcription factors in the nuclear fraction, cell-signaling receptors in the membrane fraction, soluble proteins in the cytoplasmic fraction, and more.

 

Our Compartmental Protein Extraction Kit is user-friendly and contains enough reagents for isolating total proteins from up to 5 grams of tissue or 125 million cultured cells. The kit is scalable and may be used on sample sizes as small as 0.1 g of tissue or 2.5 million cells. No scraping, no freeze-thaw cycles, and no sonication are needed. If total proteins from tissue and/or cell lysates are needed, BioChain also offers a Total Protein Extraction kit (link Total Protein Extraction kit here) to meet the demands of the researcher.

 

Applications:

  • Extraction of nuclear fractions for the study of DNA-binding proteins such as transcription factors
  • Extraction of membrane fractions for the study of membrane proteins such as cell-signaling receptors
  • Extraction of cytoplasmic fractions for the study of soluble proteins abundant in the cytosol
  • Detection of differential post-translational modifications or differential subcellular localization of target proteins
  • Enrichment of low-abundance proteins for visualization and subsequent analysis

 

Advantages and Features:

  • Versatile:Extraction from as little as 0.1 g of tissue or 2.5 million cells and up to 5 g of tissue or 125 million cells
  • Time Efficient:Manual extraction protocol isolates up to 4 compartmental fractions in under three hours
  • Cost Effective:Competitive price <$9 per extraction from 100 mg of tissue
  • Pure:Minimal cross-contamination between protein fractions

 

High Quality Compartmental Protein Extracts

Using BioChain’s CNMCs Compartmental Protein Extraction Kit results in high-quality proteins that show an absence of smearing on an SDS-PAGE gel; extraction of total proteins is also demonstrated by the range in size from 10 kd to greater than 200 kd (Figure 1, top). Minimal cross-contamination between fractions is demonstrated by Western blotting against GAPDH (cytoplasmic fraction), histone (membrane fraction), Na+/K+ ATPase (nuclear fraction), and vimentin (cytoskeletal fraction).

Figure 1. (Top) Total protein and compartmental proteins were extracted from human brain tissue using Biochain’s Total Protein Extraction Kit and CNMCS Compartmental Protein Extraction Kit. The extracts were then run using SDS-PAGE on 5 identical gels. Coomassie staining of one piece of gel indicated distinct protein patterns of respective fractions. (Bottom) PVDF membranes transferred from 4 other pieces of gel were immunoblotted against cytoplasmic marker protein GAPDH, nuclear marker protein Histone H1, membrane marker protein Na+/K+ ATPase, and cytoskeleton marker protein vimentin.

Lane 1: Total Protein                Lane 2: Cytoplasmic Protein                  Lane 3: Nuclear Protein

Lane 4: Membrane Protein                    Lane 5: Cytoskeleton Protein