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Application Note: Synthetic CRISPR - Cas9 Ribonucleoprotein (RNP)

The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system was discovered in bacteria, where it functions as an adaptive immune system against invading viral and plasmid DNA. In this system, short DNA sequences (spacers) from invading viruses are incorporated at CRISPR loci within...

July 2000 Cell Biology

A TECHNICAL APPLICATION NEWSLETTER VOL 1 July 2000 Cell Biology Isolation and Use of Mammalian Cell Nuclei by Ned Watson Sigma-Aldrich Corporation, St. Louis, MO, USA Introduction Fractionation of cells into their subcellular components has long been a central approach in cell biology. Subcellular f...

Tag Removal by Enzymatic Cleavage

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...

Purification using Glutathione Sepharose High Performance, Glutathione Sepharose 4 Fast Flow, and Glutathione Sepharose 4B

Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016 These three chromatography media are all used for the purification of GST-tagged recombinant proteins and other S-transferases or glutathione-dependent proteins. They allow mild elution conditions that preserve protei...

Purification Using GSTrap HP, GSTrap FF, and GSTrap 4B columns

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...

Lentiviral Transduction

View Other Articles | LSI Archives | Download PDF of this Article Enhancing Lentiviral Transduction Efficiency Methods for Enhancing Lentiviral Transduction Efficiency Andrea Spencer Research and Development, Biotechnology Division, Sigma-Aldrich Abstract An experiment to directly compare three meth...

July 2000 GenElute Kit

A TECHNICAL APPLICATION NEWSLETTER VOL 1 July 2000 Molecular Biology Application Notes The GenEluteTM Endotoxin-free DNA Purification Kit: Pure DNA for Endotoxin-Sensitive Applications by Fuqiang Chen, Scott Weber and Jo Anne Kerschner Sigma-Aldrich Corporation, St. Louis, MO, USA Introduction Endot...

HTS and Process Development for Capture of Recombinant Pro-insulin from E. coli using Capto MMC

Extracted from Multimodal Chromatography (PDF), GE Healthcare, 2013 This study describes the development of a robust capture method for recombinant pro-insulin. PreDictor plates and Assist software were used to determine a chromatographic medium for capture and identify promising binding and elution...

Product Directory Home

Analytical / Chromatography • Analytical Standards • Applications • Derivatization Reagents • Gas Chromatography • HPLC • LPLC/TLC/Paper Chromatography • Microbiology • Resins & Media • Sample Prep/Purification • Solid Phase Microextraction • Syringes • Titration • Vials • More.. Materials Science •...

YeastBuster™ Protein Extraction Reagent for Fast, Efficient Extraction of Proteins from Yeast

The yeast widely studied as a model eukaryotic system. The availability of the complete genome sequence, Saccharomyces cerevisiae is expression libraries, and deletion strains provide necessary tools for genetic screens and selections.1 Yeasts are capable of critical posttranslational modifications ...

Extract-N-Amp™ Blood Product Information

The Extract-N-Amp Blood PCR kits contain all of the reagents necessary to rapidly extract genomic DNA from whole blood and amplify targets of interest by PCR (Figure 1). • Novel – single-step extraction of genomic DNA to PCR • Fast – tissue to PCR in 15 minutes • Convenient – no long enzymatic diges...

Purification of Histidine-Tagged Proteins using His MultiTrap TALON

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...

DNA & RNA Purification Bacterial Genomic Miniprep Kit

For purification of genomic DNA from a variety of cultured bacteria Description Our GenElute Bacterial Genomic Kit provides a simple and convenient technique to isolate high quality DNA from both Gram negative (Figure 1) and Gram positive bacteria (Figure 2). This kit combines the advantages of a si...

Preparative Purification Using GSTPrep FF 16/10 Column

Glutathione Sepharose 4B, FF, and HP are available in lab packs, varying from 10 to 500 ml, for affinity purification of GST-tagged proteins using batch- or column-based methods. Glutathione Sepharose media are supplied preswollen in 20% ethanol. The media are used at a final slurry concentration of...

MAT-Tag Protein Expression Combines IMAC Versatility with the Sensitivity of FLAG® Detection

Combine IMAC Versatility with the Sensitivity of FLAG® Detection The MAT-Tag™ System (metal affinity tag) utilizes a small, novel, seven amino acid (HNHRHKH) sequence created for IMAC purification of recombinant MAT-Tag™ fusion proteins and the high sensitivity detection of the antibody based FLAG® ...

HTS and Optimization of a Multimodal Polishing Step in a MAb Purification Process Using Capto Adhere

Extracted from Multimodal Chromatography (PDF), GE Healthcare, 2013 This application describes the use of Capto adhere and MabSelect SuRe chromatography media to significantly reduce the level of IgG antibody aggregates in a sample using an efficient two-step method. The method resulted in high yiel...

Performing a small-scale expression screening of histidine-tagged membrane proteins from E. coli lysates

Extracted from Purifying Challenging Proteins - Principles and Methods, GE Healthcare, 2007 Buffer preparation Lysis buffer: 20 mM sodium phosphate, 100 mM NaCl, 2 mM MgCl2, 20 mM imidazole, 0.5 mM Tris (2-carboxyethyl) phosphine hydrochloride (TCEP), 5 u/ml benzonase, 1 mg/ml lysozyme, EDTA-free pr...

What is a DNA Component?

Our plasmids are composed of both a core vector backbone and a series of DNA components. Our plasmid platform is all built around the same core backbone, which means that the DNA components within each plasmids can be interchanged into each other. Below follows a brief introduction to what we consid...

Troubleshooting Purification Methods

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...

GenElute™ RNA/DNA/Protein Plus Purification Kit

Product Description Sigma’s GenElute RNA/DNA/Protein Plus Purification Kit provides a rapid method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants. The ...

Use of BugBuster® and Lysonase™ Reagents for efficient protein extraction from Gram-positive bacteria

BugBuster® Protein Extraction Reagent has become a valuable tool for extraction of proteins from E. coli without the need for mechanical disruption. It is also known that supplementing BugBuster® (as well as its derivative, PopCulture™ Reagent) with a small amount of lysozyme enhances the efficiency...

Fractional Precipitation of Affinity Chromatography Samples

Appendix 1 extracted from Affinity Chromatography Principles and Methods GE Healthcare, 2007 Fractional precipitation is frequently used at laboratory scale to remove gross impurities from small sample volumes, and occasionally used in small-scale commercial production. Precipitation techniques sepa...

CelLytic Express

Sigma-Aldrich introduces CelLytic™ Express, a non-denaturing and highly efficient protein extraction formulation for in-culture bacterial cell lysis. This proprietary, powder formulation extracts 2-3 times more protein than conventional methods such as sonication. CelLytic Express saves time by elim...

DNA & RNA Purification Blood Genomic Miniprep Kit

Description: The GenElute™ Blood Genomic DNA Kit provides a simple and convenient technique to isolate high-quality genomic DNA from fresh or aged (older than 24 hours) whole blood. This kit combines the advantages of a silica-based system with a microspin format, eliminating the need for expensive ...

Purification Using GST SpinTrap

GST SpinTrap columns are designed for rapid, small-scale purification of GST-tagged proteins using mild conditions of affinity purification. More than 90% purity can be achieved in a single step. The columns are suitable for purification of multiple samples in parallel, for screening experiments, or...

DNase I Recombinant, RNase-free Protocol

Product No. 04716728001 REMOVAL OF DNA FROM RT-PCR REACTIONS: Overview DNase I from bovine pancreas is a glycoprotein of Mr 37000. A special procedure is used to remove RNases from the DNase preparation. DNase I is a DNA-specific endonuclease that hydrolyzes ds or ssDNA to a mixture of oligo and mon...

Performing an isolation of native complexes

Related Links • Cell disruption and membrane preparation • Conditioning Purified Membrane Proteins • Performing a purification and on-column refolding of an insoluble histidine-tagged protein from a 100 ml E. coli culture using HisTrap FF 1 ml with ÄKTAprime plus • Performing a purity and homogeneity...

Histopaque® Troubleshooting Guide

Download BioFiles v6 n5 (4.55 Mb PDF) The following recommendations for troubleshooting the use of Histopaque and ACCUSPIN™ in cell separation techniques have been compiled by Sigma-Aldrich® Technical Services. These are based on observations and experience from resolving customer problems. This gui...

Validation of RNAi Knockdown Using Multiple Reaction Monitoring and Protein-AQUA™

• • Download PDF • Beth K. Radwanski (pictured), Shantanu Roychowdhury, Henry Duewel Research and Development, Biotechnology Division, MiiliporeSigma The field of proteomics is continually looking for new ways to investigate protein dynamics within complex biological samples. Recently, many research...

Chromolith® WP 300 Epoxy

Related Products • HPLC and UHPLC Columns • Chromolith® WidePore 300 HPLC Columns Related Categories • High Performance Liquid Chromatography (HPLC & UHPLC) Chromolith® WP 300 Epoxy columns are specially designed for the user-specific immobilization of ligands and their later application in HPLC. Th...