16×10/15ml Angle Rotor

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

Figure 1 / 5

Previous

Next

Click for expanded view

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

Description 

SMOBIO’s B-BOX™ is a long wavelength, blue light LED epi-illuminator. It is compact in design and robust in structure. The B-BOX™ epi-illuminator provides an unprecedented level of safety for its user due to its non-UV light source and a low operating voltage of only 12 Volts, as well as its capability working with non-carcinogenic DNA/ protein dye. The B-BOX™ comes with high quality non-flickering LED light that is highly sensitive for DNA and protein dye. B-BOX™ can greatly ease the routine chore of gel extraction, enabling easy visualization and gel cutting even in bright ambient light. The multi-angle filter plate (Cat. No. VE0102) provides optimal angles for gel cutting, visualization, and documentation. The B-BOX™ also comes with amber colored filter goggles (Cat. No. VE0103) for gel cutting and visualization. The Phox™ Photobox (Cat. No. VE0104) provides a mini darkroom environment for images taken with any smartphone. Finally, a built-in barrier system around the working area helps facilitate cleaning.

Features

• Improved cloning efficiency 
• Compact, lightweight, and portable (less than 1 kg in weight)
• Safety features include : 470 nm long wavelength, without any UV radiation hazard to its user
• Compatible with non-carcinogenic, non-ethidium bromide DNA staining dye
• User friendly: Samples are easy to visualize (when using the filter plate or goggles) 
• LED light source lasts up to 50,000 hours
• Superior detection sensitivity: ≤ 0.04 ng of DNA when using FluoroStain™ DNA Fluorescent Staining Dye, ≤3 ng of protein when using FluoroStain™ Protein Fluorescent Staining Dye (as sensitive as silver stain)
• Adjustable and removable filter plate allows for gel cutting, visualization, and documentation
• Built-in barrier design, for easy clean up
• Visible in bright ambient light
• Emphasizes minimal power reliance, low heat generation, with its own built-in heat sink

SMOBIO’s B-BOX™ is a long wavelength, blue light LED epi-illuminator. It is compact in design and robust in structure. The B-BOX™ epi-illuminator provides an unprecedented level of safety for its user due to its non-UV light source and a low operating voltage of only 12 Volts, as well as its capability working with non-carcinogenic DNA/ protein dye. The B-BOX™ comes with high quality non-flickering LED light that is highly sensitive for DNA and protein dye. B-BOX™ can greatly ease the routine chore of gel extraction, enabling easy visualization and gel cutting even in bright ambient light. The multi-angle filter plate (Cat. No. VE0102) provides optimal angles for gel cutting, visualization, and documentation. The B-BOX™ also comes with amber colored filter goggles (Cat. No. VE0103) for gel cutting and visualization. The Phox™ Photobox (Cat. No. VE0104) provides a mini darkroom environment for images taken with any smartphone. Finally, a built-in barrier system around the working area helps facilitate cleaning.

Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (<57°C)
– Pipetting aid in form of a blue dye for better visibility during aliquoting into well-plates
– included lysis function*

* = A sample extraction and sample lysis is not necessary as immediate hot PCR cycling conditions can brake cell- and virus membranes. Please see our published references to this product.

Note: Volcano3G® RT-PCR Probe 2x Master Mix is a unique master mix. If you are using it for the first time, we recommend to design primers to have higher melting/annealing temperatures (above 65°C). Additionally, for assay setup it is advised to perform RT-PCR test reactions with temperature gradients (i.e., for the annealing and extension steps) to identify the most proficient PCR protocol for your needs using our Volcano DNA polymerase products.

For research use and further manufacturing.

In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.

Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (