DBCO-PEG4-Desthiobiotin is a PEG linker containing a desthiobiotin group and a DBCO functional group. Desthiobiotin is used for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins while DBCO is a click chemistry handle that quickly reacts with azide groups on target molecules. Desthiobiotin is a sulfur-free analogue of biotin which binds streptavidin with slightly less strength than biotin, which provides it with a soft-release characteristic that is useful for in pull-down assays by minimizing co-elution with endogenous biotinylated molecules. The inclusion of a PEG linker in this molecule improves its aqueous solubility.
Detail
DBCO-PEG4-Desthiobiotin is a PEG linker containing a desthiobiotin group and a DBCO functional group. Desthiobiotin is used for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins while DBCO is a click chemistry handle that quickly reacts with azide groups on target molecules. Desthiobiotin is a sulfur-free analogue of biotin which binds streptavidin with slightly less strength than biotin, which provides it with a soft-release characteristic that is useful for in pull-down assays by minimizing co-elution with endogenous biotinylated molecules. The inclusion of a PEG linker in this molecule improves its aqueous solubility.
Other Products
ProteoSpin™ Inclusion Body Protein Isolation Kits
Product Info
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Product Info
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.
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. 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. Once opened, the solutions should be stored at 4°C when not in use except for the pH Binding Buffers (Acidic and Basic). 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
Dietary fiber can generally be described as the carbohydrate content of food that is not digested in the human small intestine. It passes into the large intestine where it is partially or fully fermented. These characteristics of dietary fiber are associated with its numerous well documented health benefits.
Dietary Fiber is a mixture of complex organic substances, including hydrophilic compounds, such as soluble and insoluble polysaccharides and non-digestable oligosaccharides, as well as a range of non-swellable, more or less hydrophobic, compounds such as cutins, suberins and lignins. The procedures for the determination and analysis of total dietary fiber as outlined in our assay protocol are based on the methods of Lee et al.1 and Prosky et al.2,3 (AOAC 991.43, AOAC 985.29, AACC 32-07.01 and AACC 32-05.01). However, the enzymes in the Megazyme Total Dietary Fiber Kit can also be used in other dietary fiber analytical methods such as AACC Method 32-21.01 and AACC Method 32-06.01.
1. Association of Official Analytical Chemists. (1985). Official Methods of Analysis, 14th ed., 1st suppl. Secs. 43, A14-43, A20, p.399. 2. Association of Official Analytical Chemists. (1986). Changes in methods. J. Assoc. Off. Anal. Chem., 69, 370. 3. Association of Official Analytical Chemists. (1987). Changes in methods. J. Assoc. Off. Anal. Chem., 70, 393.
See General Referee Reports: Journal of AOAC INTERNATIONAL, Vol. 81, No. 1, 1998.
Two separate methods are described in the associated assay protocol:
METHOD 1: DETERMINATION OF TOTAL, SOLUBLE AND INSOLUBLE DIETARY FIBER Based on AOAC Method 991.43 “Total, Soluble, and Insoluble Dietary Fiber in Foods” (First Action 1991) and AACC Method 32-07.01 “Determination of Soluble, Insoluble, and Total Dietary Fiber in Foods and Food Products” (Final Approval 10-16-91).
METHOD 2: DETERMINATION OF TOTAL DIETARY FIBER Based on AACC method 32-05.01 and AOAC Method 985.29.
Note that a letter of endorsement from the original method developer, Dr. Leon Prosky, is included in the Documents Tab.
DBCO-Maleimide is a sulfhydryl reactive reagent containing a maleimide group and a DBCO moiety. Maleimide group specifically and efficiently reacts with thiols to form thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety into cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
DBCO-Maleimide is a sulfhydryl reactive reagent containing a maleimide group and a DBCO moiety. Maleimide group specifically and efficiently reacts with thiols to form thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety into cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
