Overview

• All-in-one solution for inclusion body protein isolation and purification
• Fast and convenient spin column protocol
• Complete kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

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.

Details

Supporting Data

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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

Introduction

This product is suitable for rapid RNA extraction from tissue , cells, and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from tissue, cell
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	Tissues, cells, lymphocytes and other clinical sample
Sample amount	Cells grown in suspension:3~5 x 106Animal tissue: 10~20mgPlant tissue: ≤100mg

Principle

The Kit combines the speed and efficiency of silica-based technology with the convenient handling of magnetic particles for purification of total RNA. Samples are lysed and RNA is purified from lysates in one step through its binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet and DNA is removed by treatment with RNase-free DNase I. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water

Advantages

• High purity – OD260 / 280 :1.9-2.0, OD 260 / 230 :1.5-2.0
• Economy – less than 50% of the price of Qiagen and other imported products
• Automatic – extraction without manual participation, saving time and effort
• Universal – suitable for various clinical samples

Kit Contents

Contents	IVD3020
Purification Times	200 Preps
MagPure RNA Particles	7 ml
DNase I	4 x 600 µl
DNase Buffer	80 ml
RTL Lysis Buffer	150 ml
Buffer MCB*	75 ml
Buffer MW1*	110 ml
Buffer MW2*	50 ml
RNase Free Water	60 ml

Cat.No	Reagent	IVD3020-F-96
DNase Buffer		60 ml
DNase I		2 x 600 μl
RTL Lysis Buffer		80 ml
Buffer MCB		18 ml
96-Tip		1
Sample plate (DW Plate)	500μl Buffer MCB	1
Wash 1 Plate (DW Plate)	700μl Buffer MW130μl MagPure RNA Partilces	1
DNase Plate	Empty
Wash 2 Plate (DW Plate)	700μl Buffer MW1	1
Wash 3 Plate (DW Plate)	900μl Buffer MW2	1
Elution plate (DW Plate)	80μl RNase Free Water	1

Cat.No	Reagent	IVD3020-TL-06
Purification times		96 Preps
DNase I		2 x 600 μl
DNase Buffer		60 ml
RTL Lysis Buffer		60 ml
Buffer MCB		40 ml
96-Tip		12 PCS
2.0ml V-bottom plate	Row 1/7：500μl Buffer MCBRow 2/8：500μl Buffer MW1Row 3/9：emptyRow 4/10：30μl Magpure RNA Particles500μl Buffer MW2 Row 5/11：900μl Buffer MW2 Row 6/12：80μl RNase Free Water	6

Storage and Stability

MagPure RNA Particles should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles up to 8 weeks) at roomtemperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

This product is suitable for rapid RNA extraction from tissue , cells, and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR and so on.