Introduction

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.

Details

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

Principle

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.

Description

Specifications

Clone	IHC615
Source	Mouse Monoclonal
Positive Control	Tonsil, Follicular Lymphoma, Diffuse Large B-Cell Lymphoma
Dilution Range	1:200

LMO2, also known as LIM-Only transcription factor 2, RBTN2, or TTG2, is an oncoprotein that is expressed in normal germinal center B-cells, as well as bone marrow hematopoietic precursors and endothelial cells. LMO2 plays a role in angiogenesis and hematopoesis, and its expression has been detected in erythroid and myeloid precursors, megakaryocytes, and also in lymphoblastic and acute myeloid leukemias. LMO2 protein expression has been noted in diffuse large B-cell lymphoma, the most common adult non-Hodgkin lymphoma, as well as follicular lymphoma, a neoplasm derived from germinal center B-cells that accounts for a number of cases of non-Hodgkin lymphomas.

Overview

• Purification and enrichment of intact cell culture media exosomes for functional studies
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• Versatile cell culture media input volumes
• No phenol extractions, Proteinase K treatment, nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes required
• No precipitation reagents nor overnight incubation required
• Pure exosomes are purified and are free from any other RNA-binding proteins
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin
• The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services

Norgen’s Cell Culture Media Exosome Purification and RNA Isolation Kits constitute an all-in-one system for the purification of exosomes and the subsequent isolation of exosomal RNA from different cell culture media sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The kit is designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. 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, the 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. 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.

Cell Culture Media Exosome Purification and RNA Isolation Mini Kit

For sample input volumes ranging from 5 mL to 10 mL.

Cell Culture Media Exosome Purification and RNA Isolation Midi Kit

For sample input volumes ranging from 10 mL to 20 mL.

Cell Culture Media Exosome Purification and RNA Isolation Maxi Kit

For sample input volumes ranging from 20 mL to 35 mL.

Details

Supporting Data

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Kit Specifications
Sample Type	Cell-Free Cell Culture Media
Sample Volume Range	20 mL to 35 mL
Size of RNA Purified	All sizes, including small RNA (<200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	40 to 45 minutes
Average Yields*	Variable depending on specimen

* Please check page 4 of the product insert for Average Yields and Common RNA Quantification Methods

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. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Component	Cat. 60700 (50 preps)	Cat. 60800 (25 preps)	Cat. 60900 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	1.5 mL	1.5 mL	1.5 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	20 mL	20 mL	20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Filter Spin Column	50	25	15
Mini Spin Columns	50	25	15
Collection Tubes	50	25	15
Elution tubes (1.7 mL)	100	50	30
Product Insert	1	1	1