Overview

• Efficient depletion of bovine exosomes from Fetal Bovine Serum
• Deplete exosome-sized vesicles from versatile FBS volumes
• No protease treatment required
• No time-consuming ultracentrifugation
• No precipitation reagents required
• No overnight incubation required
• Exosome depletion confirmed by reduction of bovine miRNAs below detectable levels
• The depleted FBS provides the same cellular growth rates as the standard FBS
• Purification is based on Norgen’s proprietary Silicon Carbide resin matrix

Norgen’s FBS Exosome Depletion Kits provides a quick and easy protocol for the depletion of bovine exosomes from FBS prior to using it as a growth supplement in your culture medium. The FBS recovered from the depletion process is exosome-depleted and does not contain any quantifiable bovine miRNAs. Moreover, the exosome-depleted FBS will support the growth of your cells of interest similar to the non-depleted FBS. Norgen’s kits allow for the processing of different FBS volumes. The depletion is based on Norgen’s proprietary resin. These kits provide a clear advantage over other available kits in that they do not require ultracentrifugation, any special instrumentation, precipitation reagents or any protease treatments. More importantly, the depletion process is an inexpensive method for exosome depletion from FBS, as compared to the expensive current ready-to-use exosome-depleted media available on the market.

Background

Most culture medium used for the growth and propagation of cells in culture require the addition of fetal bovine serum (FBS) as a growth supplement to media. FBS is obtained from bovine (cow) serum, and therefore contains large quantities of bovine exosome vesicles. These exosomes may interfere with some types of studies, or may lead to unreliable results when studying the exosomes shed from your cells of interest in normal culture conditions. Therefore, the use of exosome-depleted FBS is highly recommended for many types of studies.

FBS Exosome Depletion Kit (Slurry Format)

For FBS volumes ranging from 140 mL to 280 mL.

FBS Exosome Depletion Kit (Column Format)

For FBS volumes ranging from 120 mL to 240 mL.

Details

Supporting Data

Figure 1 / 3

Previous

Next

Click for expanded view

Kit Specifications
Sample Type	Fetal Bovine Serum
Sample Volume Range	Up to 140 mL (FBS Exosome Depletion Kit I (Slurry Format) Up to 280 mL (FBS Exosome Depletion Kit II (Slurry Format)
Depletion	Deplete exosome-sized vesicle
Bovine miRNA	No detectable bovine miRNA
Time to Complete 6 Purifications	40 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. 61100 (6 preps)	Cat. 61400 (12 preps)	Cat. 61200 (6 preps)	Cat. 61300 (12 preps)
ExoC Buffer	2 x 1.5 mL	8 mL	2 x 1.5 mL	8 mL
Slurry E	12.5 mL	2 x 12.5 mL	–	–
Maxi Spin Column	–	–	6	12
Product Insert	1	1	1	1

Overview

• Sequentially purify total RNA (and miRNA), DNA and proteins from a single sample
• No sample splitting or need to use phenol or precipitation methods
• Purify RNA/DNA/Protein from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

RNA/DNA/Protein Purification Plus Kit

This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.

RNA/DNA/Protein Purification Plus Micro Kit

This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.

RNA/DNA/Protein Purification 96-Well Plus Kit

The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.

Details

Supporting Data

Figure 1 / 4

Previous

Next

Click for expanded view

Kit Specifications
Maximum Column Binding Capacity	50 μg for RNA 20 μg for DNA 200 μg for protein
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Size of DNA Purified	≥ 30 kb
Maximum Amount of Starting Material: Animal Cells Animal Tissues Blood Bacteria Yeast Fungi Plant Tissues	5 x 106 cells 25 mg (for selected tissues) 100 μL 1 x 109 cells 1 x 108 cells 50 mg 50 mg
Time to Complete 10 Purifications	30 minutes
Average Yield:HeLa Cells (1 x 106 cells)HeLa Cells (1 x 106 cells)HeLa Cells (1 x 106 cells)	15 μg RNA8 μg DNA150 μg protein

 
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. 47700 (50 preps)	Cat. 51600 (50 preps)	Cat. 51700 (96 preps)
Buffer SKP	40 mL	40 mL	–
Lysis Buffer Q	–	–	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL	20 mL
Elution Buffer F	15 mL	15 mL	2 x 15 mL
Wash Solution C	30 mL	30 mL	60 mL
Binding Buffer A	8 mL	8 mL	8 mL
Elution Buffer C	8 mL	8 mL	30 mL
Protein Neutralizer	4 mL	4 mL	4 mL
Protein Loading Dye	2 mL	2 mL	3 x 2 mL
gDNA Purification Columns	50	–	–
gDNA Purification Micro Columns	–	50	–
gDNA Purification 96-Well Plate	–	–	1
RNA/Protein Purification Columns	50	–	–
RNA/Protein Purification Micro Columns	–	50	–
RNA/Protein Purification 96-Well Plate	–	–	1
Collection Tubes	150	150	–
Collection Plate	–	–	5
Elution Tubes (1.7 mL)	150	150	–
Elution Plate	–	–	3
Lysis Preparation Plate	–	–	2
Adhesive Tape	–	–	4
Product Insert	1	1	1

Introduction

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.

Details

Specifications

Features	Specifications
Main Functions	Co-isolation DNA and RNA from skin, muscle, connective tissue, fibrous tissue sample
Applications	PCR and southern blot, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Cultured cells and tissue samples
Sample amount	Tissue: < 20mg

Principle

The Kits are designed to purify both genomic DNA and total RNA from the same cellor tissue sample. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column and bind DNA. Ethanol is added to the flow-through and the sample is applied to an RNA column. DNA/RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit. High-quality DNA is eluted in as little as 50µl water using the Kit.

Advantages

• High quality – high purity total RNA / DNA can be directly used in a variety of downstream applications
• Fast – column method can complete the extraction of several samples in 30 minutes
• Safe – no phenol chloroform extraction required
• Simultaneous extraction- simultaneously isolate DNA and RNA from one sample

Kit Contents

Contents	R511402	R511403
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RL	30 ml	150 ml
RNA Digestion Buffer	15 ml	80 ml
Buffer DW1	30 ml	150 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	2 x 50  ml
RNase Free Water	10 ml	30 ml
Buffer AE	10 ml	50 ml

Storage and Stability

HiPure DNA/RNA Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the Buffer RLC. Dissolve by warming buffer to 37°C.

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.