Gly-Gly-Gly-PEG4-DBCO

Overview

• Simple protocol that is free of detergents
• Fast processing time
• Process a variety of inputs
• Compatible with mass spectrometry and NMR spectroscopy
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a fast and simple procedure for the isolation of total proteins from tissue, bacteria, yeast or mammalian cells without the use of SDS, Triton® X-100 and other detergents. Detergents are extensively used to prepare protein samples; however, these detergents have undesirable effects on downstream analysis. These effects include extraneous peaks in mass spectrometry, artifacts with chromatography and electrophoresis, interference with microinjection into cells and interference with protein immunization. 

The Detergent-Free Total Protein Isolation Kit maintains high protein recovery and yields proteins that are 100% detergent-free.  Purification is based on using Norgen’s proprietary resin together with Lysis Solution, followed by protein filtration using a filter column (provided). The purified proteins can be used in a number of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, NMR spectroscopy and more.

Each Lysis Tube is able to process up to 50 mg of tissue, 10¹⁰ bacterial cells, 10⁹ yeast cells or 10⁷ mammalian cells. Preparation time for 12 samples is less than 10 minutes. 

Details

Supporting Data

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Kit Specifications
Maximum Amount Of Starting Material: Tissues Animal Cells Bacteria Yeast	50 mg 1 x 107 cells 1 x 1010 cells 1 x 109 cells
Time to Process 12 Samples	Less than 10 minutes

Storage Conditions and Product Stability
The Lysis Solution should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

Component	Cat. 30300 (50 preps)
Lysis Solution	4 mL
Lysis Tubes	25
Filter Columns	25
Elution Tubes (1.7 mL)	25
Product Insert	1

Introduction

Intended Usage:
For cultivating fastidious bacteria.

Principle:
Tryptone, peptone, and yeast extract multivalent powder provides a nitrogen source, vitamins, and growth factors; sodium chloride to maintain osmotic balance; glucose carbon source; dipotassium hydrogen phosphate as a buffering agent.

Formulation（per liter）:
Tryptose 20.0g
Glucose 2.0g
Dipotassium hydrogen phosphate 2.5g
Sodium chloride 5.0g
Final pH 7.3±0.2

How to use：

Suspend 29.5g in 1L of purified water. Heat with frequent agitation to dissolve the powder.
Sterilize at 121℃ for 15 minutes

Storage:

Keep the container tightly closed. Store in a cool, dry place, away from bright light. The storage period is 3 years.

Specification: 500g/bottle

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