m-PEG4-DBCO

Description 

The PM2500 ExcelBand™ 3-color Regular Range Protein Marker is a ready-to-use three-color protein standard with 10 pre-stained proteins covering a wide range of molecular weights from 10 to 180 kDa in Tris-Glycine Buffer (9 to 170 kDa in Bis-Tris (MOPS) buffer and 10 to 170 kDa Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Tris-Glycine buffer). PM2500 ExcelBand™ 3-color Regular Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.

Features

• Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
• Two reference bands — 75 kDa (red) and 25 kDa (green)

Contents

Approximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25°C), 2% SDS, 0.2 mM DTT, 3.6 M urea, and 15% (v/v) glycerol). 

Quality Control

Under suggested conditions, PM2500 ExcelBand™ 3-color Regular Range Protein Marker resolves 10 major bands in 15% SDS-PAGE (Tris-Glycine buffer, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. 

Storage

4°C for 3 months
-20°C for long term storage

The PM2500 ExcelBand™ 3-color Regular Range Protein Marker is a ready-to-use three-color protein standard with 10 pre-stained proteins covering a wide range of molecular weights from 10 to 180 kDa in Tris-Glycine Buffer (9 to 170 kDa in Bis-Tris (MOPS) buffer and 10 to 170 kDa Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Tris-Glycine buffer). PM2500 ExcelBand™ 3-color Regular Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.

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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Kit Specifications
Urine Input (Cat. 57700)	250 μL – 1 mL
Urine Input (Cat. 57800)	2 mL – 10 mL
Urine Input (Cat. 57900)	11 mL – 30 mL
Size of Exosomes Purified	40 nm – 150 nm
Elution Volume	Variable depending on the urine input volume
Time to Complete 10 Purifications	15 – 30 minutes

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.

Component	Cat. 57700 (50 preps)	Cat. 57800 (25 preps)	Cat. 57900 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Mini Filter Spin Columns inserted into 2 mL tubes	50	25	15
Product Insert	1	1	1

DBCO-NHCO-PEG4-NHS ester is a PEG linker containing NHS ester that is able to react specifically and efficiently with primary amines such as the side chain of lysine residues or aminosilane-coated surfaces at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and can also provide a long and flexible connection that minimizes steric hindrance during ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-NHCO-PEG4-NHS ester is a PEG linker containing NHS ester that is able to react specifically and efficiently with primary amines such as the side chain of lysine residues or aminosilane-coated surfaces at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and can also provide a long and flexible connection that minimizes steric hindrance during ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.