N-(t-butyl ester-PEG2)-N-bis(PEG2-propargyl)

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

This product is suitable for extracting total RNA from 50-150mg conventional plantor fungal samples as well as samples rich in polyphenolic and polysaccharides. This kit is based on silica gel column purification technology, and the extraction process does not require the use of toxic phenol chloroform extraction. The entire extraction process only takes 20-30 minutes. This kit adopts DNA filtration technology, which can efficiently filter and remove DNA. The obtained RNA can be directly used for experiments such as RT-PCR, Northern Blot, poly A+ purification, nucleic acid protection, and in vitro translation.

Details

Kit Contents

Contents	R415002	D415003
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
gDNA Filter Columns	50	250
2ml Collection Tubes	100	500
TCEP (1M)	0.29 g	5 x 0.29 g
Buffer EP	1.0 ml	5.0 ml
Buffer PSL	50 ml	250 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Except TCEP, the kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. After receiving the product, it is recommended to store TCEP (dry powder) at -20-8°C. At low temperatures, Buffer PSL may form precipitates, dissolve it completely by 55°C water bath.

Purchase Guide

1. When dealing with woody or uncommon samples, R4150 is recommended first. R4150 contains two polysaccharide/polyphenol lysis buffer, which is the most universal product.

2. R4151 is recommended for handling common economic crop samples for the first time. Strong lysis solution can be used to process easy-extraction samples. The amount of corn or rice leaves samples can reach up to 300mg.

3. R4165 adopts CTAB/chloroform method, which can also handle a large number of difficult-to-extraction plants, but requires contact with chloroform substitutes, which is less safe than other kits. This kit uses DNase Ⅰ to remove DNA, which is also a good choice for extracting polysaccharide/polyphenol-rich plant samples.

4. R4014 is recommended for fruit/starch plant samples, which uses improved trizol pre-treatment, single column operation and is more economical.

This product is suitable for extracting total RNA from 50-150mg conventional plantor fungal samples as well as samples rich in polyphenolic and polysaccharides. This kit is based on silica gel column purification technology, and the extraction process does not require the use of toxic phenol chloroform extraction. The entire extraction process only takes 20-30 minutes. This kit adopts DNA filtration technology, which can efficiently filter and remove DNA. The obtained RNA can be directly used for experiments such as RT-PCR, Northern Blot, poly A+ purification, nucleic acid protection, and in vitro translation.

• Non-specific amplification is prevented by using this Taq DNA polymerase variant as the DNA polymerase is hotstart-formulated. The 10x reaction buffer supplied together with the DNA polymerase has been specifically designed for optimal PCR performance and DNA polymerase activity. This allows the use of this DNA polymerase in a wide range of PCR applications.For research use and further manufacturing. Designed and manufactured under ISO13485. Please contact us for bulk quantities and quotes.

Broad Amplification Range

Different-sized amplicons from 3 ng of a DNA plasmid were amplified. The use of Taq DNA polymerase resulted in clean and high yield of products, as analysed after PCR on a 0.8% agarose gel.

Faster Detection and Higher Sensitivity

A fragment (64 bp) of the human blood-coagulation factor IIa (F2) was amplified from 20 ng, 2 ng, 200 pg and 20 pg of a human genomic DNA extract. The same experiment was performed in parallel using the Taq DNA polymerase mix from another well-established and known supplier. PCR products were subsequently analysed on a 2.5% agarose gel.

Non-specific amplification is prevented by using this Taq DNA polymerase variant as the DNA polymerase is hotstart-formulated. The 10x reaction buffer supplied together with the DNA polymerase has been specifically designed for optimal PCR performance and DNA polymerase activity. This allows the use of this DNA polymerase in a wide range of PCR applications.