DBCO STP ester

Overview

• Sequentially purify total RNA and total proteins from a single sample
• Kit includes a gDNA elimination column  
• No sample splitting required
• No phenol step required for efficient isolation
• Ideal for small or difficult to obtain samples
• Purify RNA and proteins from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Rapid and efficient spin column procedure
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.

Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.  

This kit eliminates DNA efficiently using a gDNA removal column.

Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA.  The proteins will not require precipitation, resuspending of pellets, or any further cleaning.

This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are 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 to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.

Protocol

Details

Supporting Data

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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
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). 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. 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

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• ​Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment, nor carrier RNA
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents, nor overnight incubation required
• Concentrate isolated exosomal RNA and are free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 50 µL to 1 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 1 mL to 4 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 4 mL to 10 mL.

Details

Supporting Data

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Kit Specifications
Minimum Plasma Input	1 mL
Maximum Plasma Input	4 mL
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50 – 100 µL
Time to Complete 10 Purifications	40 – 45 minutes
Average Yields*	Variable depending on specimen

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Component	Cat. 59500 (50 preps)	Cat. 59600 (25 preps)	Cat. 59700 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1