Overview

• Columns bind proteins of interest while endotoxins flow through
• Reduce endotoxin levels to less than 0.01 EU/µg of protein
• Proteins are desalted
• Greater than 95% protein recovery
• Concentrate protein samples and remove detergents at the same time
• Effectively remove a wide range of detergents including SDS, Triton® X-100, CHAPS, NP-40, and Tween 20
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Kits are designed for the rapid removal of endotoxins from previously purified proteins or peptides, with protein recoveries of > 95% being achieved. Endotoxins, also known as lipopolysaccharides, are cell-membrane components of Gram-negative bacteria such as E. coli. Endotoxins liberated by Gram-negative bacteria are frequent contaminations of protein solutions derived from bioprocesses. Due to the high toxicity of endotoxins in vivo and in vitro, their removal from protein preparations is often necessary prior to the use of the protein in downstream applications. These kits efficiently reduce endotoxin levels to ≤ 0.01 EU/mg of protein, using spin column chromatography based on Norgen’s proprietary resin as the separation matrix. These kits are highly efficient in removing many different salts commonly used in the laboratory including, but not limited to, MgCl₂, NaCl, KCl, CaCl₂, LiCl and CsCl. The purified protein samples can be used in a number of downstream applications including sequencing, cloning, and in vitro and in vivo introduction into cells and organisms for various purposes. The simultaneous removal of salts while concentrating a dilute protein solution makes the kit a convenient method for preparing proteins before running many downstream applications such as SDS-PAGE, isoelectric focusing, X-ray crystallography, NMR spectroscopy, mass spectroscopy and other applications.

Mini Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Mini Kit is designed for the rapid removal of endotoxins from up to 200 μg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Maxi Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Maxi Kit is designed for the rapid removal of endotoxins from up to 4 mg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Details

Supporting Data

Figure 1 / 15

Previous

Next

Click for expanded view

Kit Specifications
Maximum Protein Input	200 μg
Maximum Column Volume Input	600 μL
Molecular Weight of Recovered Proteins	No Molecular Weight cut-off
Final Endotoxin Levels	≤ 0.01 EU/μg protein
Protein Recovery	90-95%
% Detergent Removal	90-95%
Detergents that can be Removed	Including Triton® X-100, CHAPS, NP-40 and Tween 20
Minimum Elution Volume	50 μL
Time to Complete 10 Purifications	20 minutes

Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.

Component	Cat. 22800 (25 preps)	Cat. 22200 (4 preps)
Binding Buffer J	8 mL	8 mL
Binding Buffer N	4 mL	20 mL
Wash Solution M	50 mL	130 mL
Wash Solution CIP	20 mL	60 mL
Wash Solution N	30 mL	130 mL
Wash Solution NIP	20 mL	60 mL
Elution Buffer G	6 mL	20 mL
Endotoxin Removal Solution	1.5 mL	1.5 mL
Protein Neutralizer EF	2 mL	2 mL
Maxi Spin Columns (assembled with Collection Tubes)	–	4
Mini Spin Columns	25	–
Collection Tubes	25	–
Maxi Spin Columns (assembled with Collection Tubes)	4
Elution Tubes (1.7 mL)	25	–
Elution Tubes (50 mL)	4	–
Product Insert	1	1

Introduction

This product provides a fast and easy way to purify DNA from plant and fungal tissue. Up to 100 mg of tissue can be processed. Easy-to-use Plant procedures provide pure total DNA (genomic,mitochondrial, and chloroplast) for reliable PCR and southern blotting in less than 1 hour. Purification requires no phenol or chloroform extraction oralcohol precipitation and involves minimal handling.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from100 mg simple plant
Applications	PCR, SSR, AFLP, RAPD and southern blot, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Economic plant samples
Sample amount	Fresh / frozen plant samples: ≤100mggDried plant / seed samples: ≤20mg
Elution volume	≥30μl
Time per run	30-50 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

Plantmaterial is first mechanically disrupted and then lysed by addition of lysis buffer and incubation. RNase A in the lysis buffer digests the RNA in the sample. After lysis, proteins and polysaccharides are salt-precipitated.Binding buffer and ethanol are added to the cleared lysate to promote binding of the DNA to the HiPure membrane. The sample is then applied to a column and then centrifuged. DNA binds to the membrane, while contaminants such as proteins and polysaccharides are efficiently removed by 2 wash steps. Pure DNA is eluted in a small volume of low-salt buffer or water.

Advantages

• Safe – require no phenol or chloroform extraction
• Fast – several samples can be extracted in 30 minutes by silica technology
• High purity – high quality DNA, completely remove inhibitors
• High yield – silica technology can achieve the highest yield

Kit Contents

Contents	D316402	D316403
Purification Times	50 Preps	250 Preps
RNase A	10 mg	50 mg
Protease Dissolve Buffer	1.8 ml	5 ml
Buffer SPL	30 ml	150 ml
Buffer PS	10 ml	50 ml
Buffer GW1	22 ml	110 ml
Buffer GW2*	12 ml	2 x 50 ml
Buffer AE	15 ml	60 ml
HiPure gDNA Mini Columns	50	2 x 125
2 ml Collection Tubes	50	2 x 125

Storage and Stability

RNase A should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

Experiment Data

Purchase Guide

Name	CAT NO	Fresh / frozen tissue amount	Dried tissue amount	Column type	Elution volume	Yield (muscle)
HiPure Plant DNA Mini Kit	D3187	150mg	40mg	1.5ml column	50 – 100μl	3-70μg
HiPure HP Plant DNA Maxi Kit	D3163	5g	1g	50ml column	0.7 – 3ml	75-1250μg
HiPure SF Plant DNA Kit	D3164	100mg	20mg	1.5ml column	50 – 100μl	3-50μg
HiPure SF Plant DNA 96 Kit	D3167	50mg	15mg	96 well plate	75 – 150μl	3-30μg

This product provides a fast and easy way to purify DNA from plant and fungal tissue. Up to 100 mg of tissue can be processed. Easy-to-use Plant procedures provide pure total DNA (genomic,mitochondrial, and chloroplast) for reliable PCR and southern blotting in less than 1 hour. Purification requires no phenol or chloroform extraction oralcohol precipitation and involves minimal handling.