DBCO-NHCO-PEG5-acid

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

Overview

• Isolate genomic DNA from animal tissues, cells, bodily fluids, viruses and swabs 
• Rapid and convenient spin column procedure 
• Purified DNA is of the highest quality and integrity for sensitive downstream applications including PCR, qPCR, genotyping, sequencing and more

This kit is designed for the rapid preparation of genomic DNA from various tissue samples, cultured cells, viruses, bodily fluids and swabs using a rapid spin column protocol. Purified DNA is of an excellent yield and quality, and is immediately ready for any downstream application including PCR, qPCR, genotyping, sequencing and more.  The protocol can be completed in approximately 80 minutes (including incubation time).

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	25 µg
Average Yield:* HeLa Cells (1 x 106) Tissue (from 10 mg kidney)	8 µg 10 µg
Maximum Amount of Starting Material: Animal Tissues Cultured Cells Bodily Fluids (blood, saliva) Viral Suspension	20 mg 3 x 106 cells 150 µL 150 µL
Time to Complete 10 Purifications	80 minutes

* Yield will vary depending on the type of sample processed

Storage Conditions and Product Stability
The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other 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. 24700 (50 preps)	Cat. 24750 (100 preps)	Cat. 24770 (250 preps)
Digestion Buffer A	25 mL	2 x 25 mL	5 x 25 mL
Buffer SK	30 mL	2 x 30 mL	5 x 30 mL
Wash Solution A	18 mL	2 x 18 mL	5 x 18 mL
Elution Buffer B	30 mL	2 x 30 mL	5 x 30 mL
Proteinase K	12 mg	2 x 12 mg	5 x 12 mg
Spin Columns	50	100	250
Collection Tubes	50	100	250
Elution Tubes (1.7 mL)	50	100	250
Product Insert	1	1	1

Overview

Optically clear adhesive film with pressure activated adhesive. Seal is peelable; suitable for qPCR and other imaging techniques including crystallization.

• Optically clear seal specifically developed for optical applications, particularly qPCR
• It is non sticky when removed from the packaging; this aids handling when wearing gloves
• The adhesive is contained within small capsules, allowing light to pass through to ensure the optical clarity of the seal
• When the seal is in position, pressure can be applied to burst the capsules, releasing a strong adhesive only where the seal touches the raised well rims of the plate – the rest of the seal area above the wells remains optically clear
• For all adhesive seals, the best sealing results are achieved using our Hand Roller or KAPS 500 Auto Sealer

Optically clear adhesive film with pressure activated adhesive. Seal is peelable; suitable for qPCR and other imaging techniques including crystallization.