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	4 mg
Minimum Protein Input	0.25 mg
Maximum Column Volume Input	18 mL
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
Elution Volume	2-4 mL
Time to Complete 10 Purifications	30-40 minutes

Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after 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 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.

Description

Specifications

Clone	IHC412
Source	Mouse Monoclonal
Positive Control	Colon
Dilution Range	1:200

Postmeiotic Segregation Increased 2 (PMS2) is a DNA repair protein involved in mismatch repair. Mutations and deficiencies in the PMS2 gene have been linked to microsatellite instability, and malignancies such as hereditary nonpolyposis colorectal cancer and endometrial cancer. Expression levels of the PMS2 protein may be useful as a screening tool for Lynch syndrome after a colorectal cancer diagnosis. Anti-PMS2 is recommended to be used as part of a panel along with antibodies against MLH1, MSH2, and MSH6.