Overview

• Purification and enrichment of intact exosomes from plasma, serum, urine, cell culture media and saliva in less than 30 minutes.
• Versatile sample input ranging from 250 µL to 30 mL
  • Urine Exosome Purification Mini Kit (250 µL – 1 mL urine)
  • Urine Exosome Purification Midi Kit (2 mL – 10 mL urine)
  • Urine Exosome Purification Maxi Kit (11 mL – 30 mL urine)
• Exosome purification is based on Norgen’s proprietary resin separating matrix through exosomes’ surface proteins.
• No precipitation reagents, overnight incubation, protease or coagulant treatments required
• No time-consuming ultracentrifugation, filtration or special syringes required
• Purify intact exosomes with a size ranging from 40-200 nm depending on sample input type
• Purified exosomes are compatible with functional studies.
• Purified exosomes are free from any protein-bound cell-free circulating RNA
• Purified exosomes are compatible with NanoSight® or Electron Microscopy for assessing the approximate exosome size range and concentration.
• Exosomal RNA can be extracted from the purified exosomes using Norgen’s Exosomal RNA Purification technology or any other RNA extraction method.

The Urine Exosome Purification Kits provide a fast, reliable and convenient method to purify and enrich for pure intact exosomes from different urine sample volumes ranging from 250 µL to 30 mL. These kits also allow for the purification of intact extracellular vesicles (EVs) from different urine sample volumes, and these EVs are ready for any downstream application. The purification is based on Norgen’s proprietary resin.

These kits provide a clear advantage over other available methods since they do not require any special instrumentation, ultracentrifugation, precipitation reagents or any protease treatments. More importantly, the purified exosomes will not be contaminated with any other RNA-binding proteins that may contaminate your exosomal RNA, which is essential if studying exosomal RNA gene expression.

NanoSight® Analysis

Exosomes enriched with Norgen’s Urine Exosome Purification Kits can be analyzed using NanoSight® for assessing the approximate exosome size range and concentration.

Exosomal RNA Analysis

Exosomal RNA can be isolated using Norgen’s Exosomal RNA Isolation Kit from exosomes enriched using Norgen’s Urine Exosome Purification Kits for gene expression analysis using RT-qPCR, microarray or NGS and for Biomarker discovery.

Details

Supporting Data

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Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Introduction

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

This method is suitable for small quantities of tissue (<100mg) and cells (<5 X10⁶), and large quantities of tissue (up to 1g) and cells (<10⁸), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.

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Specifications

Advantages

• Flexible – sample amount can be adjusted according to the demand
• Cost performance -the most economical nucleic acid extraction technology

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

Overview

• Columns bind proteins of interest while endotoxins flow through
• Reduce endotoxin levels to less than 0.01 EU/µg of protein
• Proteins are desalted
• Greater than 95% protein recovery
• Concentrate protein samples and remove detergents at the same time
• Effectively remove a wide range of detergents including SDS, Triton® X-100, CHAPS, NP-40, and Tween 20
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Kits are designed for the rapid removal of endotoxins from previously purified proteins or peptides, with protein recoveries of > 95% being achieved. Endotoxins, also known as lipopolysaccharides, are cell-membrane components of Gram-negative bacteria such as E. coli. Endotoxins liberated by Gram-negative bacteria are frequent contaminations of protein solutions derived from bioprocesses. Due to the high toxicity of endotoxins in vivo and in vitro, their removal from protein preparations is often necessary prior to the use of the protein in downstream applications. These kits efficiently reduce endotoxin levels to ≤ 0.01 EU/mg of protein, using spin column chromatography based on Norgen’s proprietary resin as the separation matrix. These kits are highly efficient in removing many different salts commonly used in the laboratory including, but not limited to, MgCl₂, NaCl, KCl, CaCl₂, LiCl and CsCl. The purified protein samples can be used in a number of downstream applications including sequencing, cloning, and in vitro and in vivo introduction into cells and organisms for various purposes. The simultaneous removal of salts while concentrating a dilute protein solution makes the kit a convenient method for preparing proteins before running many downstream applications such as SDS-PAGE, isoelectric focusing, X-ray crystallography, NMR spectroscopy, mass spectroscopy and other applications.

Mini Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Mini Kit is designed for the rapid removal of endotoxins from up to 200 μg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Maxi Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Maxi Kit is designed for the rapid removal of endotoxins from up to 4 mg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Details

Supporting Data

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Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.