This kit provides a rapid spin column procedure for the isolation of exosomal RNA from urine samples. Users can simultaneously concentrate and isolate high quality exosomal RNA, including microRNA, for use in sensitive downstream assays such as RT-PCR, qRT-PCR, NGS, microarrays and more. The protocol can be completed in under 50 minutes. Urine volumes of 1 to 10 mL can be processed easily and rapidly. All sizes of RNA are recovered at an equal rate without the need for using hazardous chemicals like phenol.
Background
Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascite fluids, among other biological fluids. These vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which can inform about the cell type from which the exosomes are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNases they can be efficiently recovered from biological fluids, such as urine.
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| Kit Specifications | |
| Minimum Urine Input | 1 mL |
| Maximum Urine Input | 10 mL |
| Size of RNA Purified | Small exosomal RNA species |
| Time to Complete Purification | ~ 50 minutes |
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 47200 (50 preps) |
|---|---|
| Slurry B1 | 18 mL |
| Binding Buffer A | 20 mL |
| Lysis Buffer A | 2 x 20 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Mini Filter Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
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.
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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 |
Norgen’s Saliva RNA Collection and Preservation Devices are designed for simple and non-invasive saliva collection and preservation of RNA in saliva samples at ambient temperature. Each of the 50 Saliva RNA Collection and Preservation Devices consists of 3 components:
Saliva samples are collected by spitting inside the Collection Funnel which has been assembled with the Collection Tube. After collecting the required volume of saliva the Collection Funnel is removed and the contents of the Preservative Ampoule are then added and mixed with the collected saliva. The Saliva Collection Tube is subsequently sent to the laboratory for RNA isolation and analysis. RNA can be isolated from the preserved saliva samples using Norgen’s Saliva/Swab RNA Purification Kits (Cat# 69100, 69300). Each of Norgen’s Collection Tubes is labeled with a unique serial number that can be used for secure and anonymous tracking of the sample. The Saliva RNA in preserved samples is stable for up to 2 months at room temperature. This kit is ideal for collecting and preserving RNA samples for epidemiological and population studies.
Norgen’s Saliva RNA Preservative is an aqueous storage buffer designed for rapid cellular lysis and subsequent preservation of RNA from fresh specimens. The buffer eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperature. The components of the buffer allow samples to be stored for up to 2 months at room temperature.
Saliva RNA can be isolated from the preserved saliva samples using Norgen’s Saliva/Swab RNA Purification Kits (Cat# 69100, 69300).
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| Kit Specifications | |
| Volume of Saliva Collected | 2 mL |
| Volume of Saliva-Preservative Mix | 4 mL |
| Preservation Temperature | Room Temperature |
| Preservation time | Up to 2 months at room temperature |
Shelf Life and Handling
| Kit Components | Cat. RU53800 (50 Devices) |
|---|---|
| Individual Saliva RNA Collection and Preservation Devices | 50 |
| Donor Procedure Flowchart | 1 |
| Product Insert | 1 |
| Individual Saliva RNA Collection and Preservation Device Contents | |
| Saliva Collection Funnel and Collection Tube | 1 |
| Collection Tube Cap | 1 |
| Preservative Ampoule | 1 |
| Donor Instructions | 1 |