The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA and miRNA from 0.2-0.6ml serum and plasma |
| Applications | |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | |
| Sample amount | |
| Yield | |
| Elution volume | |
| Time per run | |
| Liquid carrying volume per column | |
| Binding yield of column |
HiPure RNA technology simplifies total RNA isolation. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After the removal of protein the binding conditions for nucleic acids are adjusted by adding isopropanol. Total nucleic acids are bound to the column. Optionally, DNA can be removed by an on-column DNase digest. The remaining nucleic acids are washed and eluted with minimal amounts of RNase-Free water.
Kit Contents
| Contents | R431402 | R431403 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 100 | 2 x 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| Buffer CFL | 6 ml | 30 ml |
| Buffer CFP | 1.8 ml | 10 ml |
| Buffer RWC | 20 ml | 80 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 60 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.
The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
This kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, bacterial cells, plants, and blood.
Two columns are provided with this kit. The first column captures large RNA, while the small RNA are captured on a second column and are eluted concentrated in 25 µL of nuclease-free water.
The small RNA can be used in various downstream applications relating to miRNA profiling, gene regulation, and functional analysis. The eluted RNA is ready for RT-qPCR, microarrays, and NGS applications.
Background
These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation.
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| Kit Specifications | |
| Maximum Column Binding Capacity | Up to 50 μg RNA |
| Maximum Column Loading Volume | 650 μL |
| Minimum Elution Volume | 20 μL |
| Size of RNA Purified | All sizes, including small RNA < 200 nt |
| Time to Complete 10 Purifications | 25 minutes |
| Maximum Amount of Starting Material: Animal Cells Animal Tissues Bacteria Plant Tissues Blood | 3 x 106 cells 5-25 mg 1 x 109 cells 50 mg 100 μL |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 21300 (25 preps) |
|---|---|
| Buffer RL | 40 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Large RNA Removal Column | 25 |
| microRNA Enrichment Column | 25 |
| Collection Tube | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
These kits provide everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting. This kit provides a convenient way to screen recombinants prior to scaling up.
ProteoSpin™ Inclusion Body Protein Isolation Micro Kit
The process is efficient and streamlined and can process up to 12 samples in only 60 minutes. Each spin column is able to recover up to 50 µg of acidic or basic proteins. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit
The procedure is efficient and streamlined and can process up to 4 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
About Inclusion Bodies
Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are typically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these subcellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time-consuming and not cost-effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography – all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.
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| Kit Specifications | |
| Maximum Culture Volume | 100 mL |
| Yield from 100 mL of Culture | Up to 12 mg |
| Minimum Elution Volume | 4 mL |
| Time to Process 1 Sample | 1 hour |
Storage Conditions
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. This kit is stable for 2 years after the date of shipment. Once opened, the solutions should be stored at 4°C when not in use except for Binding Buffer C and Binding Buffer N. Some precipitation will occur with 4°C storage. This precipitation should be dissolved with slight heating to room temperature before using.
| Component | Cat. 10300 (Micro – 25 preps) | Cat. 17700 (Maxi – 4 preps) |
|---|---|---|
| Wash Solution C | 30 mL | 130 mL |
| Wash Solution N | 30 mL | 130 mL |
| Binding BUffer A | 4 mL | 20 mL |
| Binding Buffer N | 4 mL | 20 mL |
| Elution Buffer C | 8 mL | 2 x 30 mL |
| Protein Neutralizer | 4 mL | 4 mL |
| Cell Lysis Reagent | 15 mL | 110 mL |
| IB Solubilization Reagent | 2 mL | 50 mL |
| Syringes, 1cc, slip tip | 25 | – |
| Needles (Bev, 20G x 1 inch) | 25 | – |
| Syringes, 10 mL, Luer-Lok™ Tip | – | 4 |
| Needles (18G x 1.5 inch) | – | 4 |
| Micro Spin Columns | 25 | – |
| Maxi Spin Columns (filled with SiC) inserted into 50 mL collection tubes | – | 4 |
| Collection Tubes | 25 | – |
| Elution Tubes (1.7 mL) | 25 | – |
| Elution Tubes (50 mL) | – | 4 |
| Product Insert | 1 | 1 |