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

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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

HiPure PCR Pure Mini Kit uses proprietary chemistry and HiPure technology to recover DNA Fragments between 70bp-30kbp with yields exceeding 85%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.

Compared with the gel purification kit, this product uses guanidine hydrochloride mediated adsorption, which have better A260/A230 ratio.

Details

Specifications

Features	Specifications
Main Functions	Recover DNA fragments >100bp from various pcr products, enzymatic reaction solutions, labeling reaction solutions, crude DNA, etc.
Applications	PCR, sequencing, labeling reactions, ligations and restriction digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	PCR products, enzymatic reaction solution
Sample amount	10-200μl
Recovery	≥85%
Elution volume	≥15μl
Time per run	≤10 minutes
Liquid carrying volume per column	800µl
Binding yield of column	35µg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.

Advantages

• High recovery efficiency – ≥85% DNA recovery
• Fast – isolation can be completed in 10 minutes by column gel method

Kit Contents

Contents	D212102	D212103
Purification Times	100 Preps	250 Preps
Buffer DP	60 ml	120 ml
Buffer DW2	20 ml	50 ml
Elution Buffer	15 ml	30 ml
HiPure DNA Mini Columns II	100	250
2 ml Collection Tubes	100	250

Storage and Stability

The Kit should be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.

Experiment Data

HiPure PCR Pure Mini Kit uses proprietary chemistry and HiPure technology to recover DNA Fragments between 70bp-30kbp with yields exceeding 85%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.

Compared with the gel purification kit, this product uses guanidine hydrochloride mediated adsorption, which have better A260/A230 ratio.

For scaling up and fully automating end-to-end NGS library prep

With the Flex NGS Workstation, you can automate your NGS workflow at the scale you need to increase efficiency, reduce errors, and save hands-on time. The Flex enables you to automate your library prep and pre-sequencing workflows using any leading reagent system on the market, including fragmentation and tagmentation protocols.

Optional add-ons can be purchased at a 10% discount when ordered with the Flex NGS Workstation*

For scaling up and fully automating end-to-end NGS library prep

With the Flex NGS Workstation, you can automate your NGS workflow at the scale you need to increase efficiency, reduce errors, and save hands-on time. The Flex enables you to automate your library prep and pre-sequencing workflows using any leading reagent system on the market, including fragmentation and tagmentation protocols.

Optional add-ons can be purchased at a 10% discount when ordered with the Flex NGS Workstation*