Sequentially isolate total RNA and proteins from the same sample for streamlined analysis.
Format
Plus
Size
50 Preps
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Features and benefits
- Purify the total proteins and RNA from a single sample in a sequential manner
- A gDNA elimination column is included in the kit
- Sample splitting is not necessary
- Phenol step is not necessary for effective isolation
- Perfect for tiny or challenging-to-get samples
- Remove RNA and proteins from tissues, blood, bacteria, yeast, fungi, or plants that have been cultured.
- Quick and effective spin column process
- Purification is based on spin column chromatography, which uses Norgen's exclusive resin separation matrix.
This kit offers a fast and efficient single-column method for sequentially isolating and purifying total RNA (including miRNA) and proteins from a single sample. It is compatible with cultured animal cells, tissues, blood, bacteria, yeast, fungi, and plants. Both total RNA and proteins are column-purified in under 25 minutes using a single column.
High-quality and high-yield purified RNA is appropriate for NGS, RT-qPCR, and microarrays.
This kit uses a gDNA removal column to remove DNA effectively.
After being eluted in buffer, proteins are prepared for use in subsequent procedures like ELISA, Mass Spec, and Western Blots. Precipitation, pellet resuspension, or additional cleaning will not be necessary for the proteins.
This kit is ideal for researchers studying the transcriptome and proteome from a single sample, making it suitable for applications such as microRNA profiling, gene expression analysis (including gene silencing and mRNA knockdown studies), biomarker discovery, and cultured cell line characterization.
Norgen’s RNA/Protein Purification Plus Kit is particularly beneficial for isolating macromolecules from precious, difficult-to-obtain, or small samples, such as biopsy materials or single foci from cell cultures, as it eliminates the need for sample fractionation. Because RNA and proteins are isolated from the same sample, the results are more reliable and consistent. The purified macromolecules are of the highest purity and suitable for a wide range of downstream applications.
Protocol
Image Credit: Norgen Biotek Corporation
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Details
Supporting data
Figure 1. Recovery of True Total RNA, including microRNA from Hamster Liver. Panel A is a 1X MOPS 1% agarose gel showing the RNA that was isolated from 2 different samples of 10 mg hamster liver using Norgen's RNA/Protein Purification Plus Kit. Norgen's RNA/Protein Purification Plus Kit isolated both large RNA (represented by 28S and 18S rRNA) as well as small RNA with high integrity and without having to perform any additional protocol. Panel B is a result from a bioanalyzer resolution of the eluted RNA. Similar to the agarose gel, the Bioanalyzer showed that Norgen's RNA/Protein Purification Plus Kit has the added benefit of recovering small RNA. One microgram of the RNA was used in RT-qPCR reactions for the detection of human beta-Actin (for Large RNA) and miR-21 (for microRNA) genes. The RNA isolated using Norgen's RNA/Protein Purification Plus Kit showed superior recovery of both large RNA and small RNA including microRNAs as depicted by the successful miR-21 RT-qPCR (Panel C). Image Credit: Norgen Biotek Corporation
Figure 2. High-Quality Total Proteins Eluted in Mass Spec-Compatible Buffer. Norgen's RNA/Protein Purification Plus Kit provides a column purification step for effective concentration and clean-up of the isolated proteins. The proteins are eluted into a buffer which is compatible with many downstream applications including mass spectrometry as well as standard protein quantification methods (including Bradford assays). In contrast, most competing multiple analyte isolation products require protein precipitation and the precipitated proteins are required to be resuspended in buffer with high-detergent content (such as SDS-PAGE loading dye) for full recovery. In this figure, the protein fraction isolated from hamster liver using Norgen's RNA/Protein Purification Plus Kit was resolved direclty on a 12% SDS-PAGE protein gel. Norgen's RNA/Protein Purification Plus Kit purified the proteins by column and the eluted proteins are already in buffer compatible with downstream applications. Image Credit: Norgen Biotek Corporation
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Kit specifications
Source: Norgen Biotek Corporation
| Kit Specifications |
| Maximum Column Binding Capacity |
50 μg for RNA |
| Maximum Column Binding Capacity |
200 μg for Protein |
| Maximum Column Loading Volume |
650 μL |
| Size of RNA Purified |
All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications |
30 minutes |
Average Yields*:
HEK 293 Cells (1 x 106 cells)
HEK 293 Cells (1 x 106 cells)
Liver (15 mg)
Liver (15 mg) |
10-15 μg RNA
70-100 μg protein
30-35 μg RNA
100-150 μg protein |
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
Storage conditions and product stability
After adding DL-Dithiothreitol (DTT), the Protein Loading Dye should be kept at -20 °C. Every solution needs to be stored at room temperature and kept tightly sealed. For a year following the date of shipment, this kit remains stable.
Source: Norgen Biotek Corporation
| Component |
Cat. 48200 (50 preps) |
| Buffer SKP |
40 mL |
| Wash Solution A |
38 mL |
| Elution Solution A |
6 mL |
| Wash Solution C |
30 mL |
| Binding Buffer A |
8 mL |
| Elution Buffer C |
4 mL |
| Protein Neutralizer |
4 mL |
| Protein Loading Dye |
2 mL |
| gDNA Removal Columns |
50 |
| RNA/Protein Purification Columns |
50 |
| Collection Tubes |
150 |
| Elution Tubes (1.7 mL) |
100 |
| Product Insert |
1 |