Overview

• Simultaneous concentration, desalting and buffer exchange of total urinary proteins
• No molecular weight cutoff allows for isolation of all sizes of proteins and peptides
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

These kits concentrate urine proteins while simultaneously removing salts, urea, and other urine contaminants. There is no molecular weight cut-off and therefore the columns capture total urinary proteins and peptides of all sizes making them ideal for biomarker discovery work, differential expression of proteins in various diseases, or other diagnostic research. The columns are convenient, rapid and easy to use and thus offer significant time savings over classic dialysis protocols. The resulting high-quality protein sample is concentrated and free from the original sample salts, thus preparing the sample conveniently for downstream proteomic applications including SDS-PAGE, 2D Gels, MALDI-TOF, LC/MS, LC/MS/MS, whole protein mass spectrometry, western blotting, protein microarrays, and more.

ProteoSpin™ Urine Protein Concentration Micro Kit

Each spin column is able to concentrate and desalt up to 200 μg of urine proteins. Twelve samples can be processed in 20 minutes.

ProteoSpin™ Urine Protein Concentration Midi Kit

The ProteoSpin™ Urine Protein Concentration Midi Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from 1 to 5 mL inputs of urine. Each mini spin column is able to concentrate and desalt up to 3 mg of urine proteins in 30 minutes.

ProteoSpin™ Urine Protein Concentration Maxi Kit

The ProteoSpin™ Urine Protein Concentration Maxi Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from 2 to 20 mL inputs of urine. Each maxi spin column is able to concentrate and desalt up to 4 mg of urine proteins in 45 minutes.

Details

Supporting Data

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Kit Specifications
Minimum Urine Volume Input	1 mL
Maximum Urine Volume Input	5 mL
Maximum Recovered Proteins (5 mL Urine)	3 mg*
Elution Volume	200 – 1000 μL
Time to Complete 10 Purifications	30 minutes

 *Yield will depend on the health status of the donor.

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

Component	Cat. 17400 (25 preps)	Cat. 52300 (10 preps)	Cat. 21600 (4 preps)
Wash Solution C	60 mL	60 mL	130 mL
Binding Buffer A	4 mL	4 mL	8 mL
Elution Buffer C	8 mL	30 mL	30 mL
Protein Neutralizer	4 mL	4 mL	4 mL
Micro Spin Columns	25	–	–
Midi Spin Columns (assembled with collection tubes)	–	10	–
Maxi Spin Columns (assembled with collection tubes)	–	–	4
Collection Tubes	25	–	–
Elution Tubes (1.7 mL)	25	–	–
Midi Elution Tubes	–	10	–
Elution Tubes (50 mL)	–	–	4
Product Insert	1	1	1

Introduction

This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.

Details

Specifications

Features	Specifications
Main Functions	Isolation both Circulating DNA/RNA (include miRNA) from 1-5 ml serum and plasma
Applications	qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection
Purification method	Midi spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	serum, plasma, and other cell-free liquid samples
Sample amount	1-5 ml
Elution volume	≥20μl
Time per run	≤100 minutes
Liquid carrying volume per column	4 ml
Binding yield of column	1 mg

Principle

This kit is based on silica gel column technology. Serum or other liquid samples are lysed and digested in buffer CFL. After adding buffer CFP, the protein is removed by centrifugation to obtain the supernatant. Isopropanol is added to precipitate the total nucleic acid and transferred to the column for filtration. DNA / RNA is adsorbed on the membrane of the column, while the protein is not adsorbed and removed with the filtrate.The column is washed with buffer MGW1 to remove protein and other impurities, and then washed with buffer RW2 to remove salt. Finally, DNA / RNA is eluted by low salt buffer. The eluted DNA / RNA can be directly used for quantitative PCR/ RT-PCR, liquid or solid-phase chip analysis, hybridization and SNP detection.

Advantages

• High yield – most optimized process to obtain maximum free RNA and small RNA
• High concentration – low elution volume (>20μl) to ensure nucleic acid concentration
• High purity – low alcohol combination, completely remove inhibitor and protein pollution
• High recovery – silica gel column technology can recover nucleic acid molecules at the level of PG
• Large volume – 1-2ml serum and plasma samples can be processed at one time

Kit Contents

Contents	R431602	D431603
Purification Times	50 Preps	250 Preps
HiPure RNA Micro Columns	50	5 x 50
HiPure Viral Midi Columns	50	5 x 50
15 ml Collection Tubes	50	5 x 50
2ml Collection Tubes	50	5 x 50
Buffer CFL	150 ml	2 x 375 ml
Buffer CFP	30 ml	150 ml
Buffer MGW1*	100 ml	2 x 250 ml
Buffer RW2*	2 x 50 ml	5 x 100 ml
RNase Free Water	10 ml	50 ml

Storage and Stability

The kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.