Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:
> 2 years under recommended storage conditions
Analyte:
Pullulanase/Limit-Dextrinase
Assay Format:
Spectrophotometer
Detection Method:
Absorbance
Wavelength (nm):
400
Signal Response:
Increase
Limit of Detection:
0.18 U/mL for pullulanase preparations (50-fold dilution) 0.01 U/g for limit dextrinase in milled malt
Reproducibility (%):
~ 3%
Total Assay Time:
~ 10 min (Pullanase), ~ 30 min (Limit-Dextrinase)
Application examples:
Assay of microbial pullulanase preparations. Measurement of limit-dextrinase in malt extracts.
Method recognition:
Novel method
PullG6 assay for the measurement of pullulanase employs a water soluble defined substrate, namely 4,6-O-benzylidene-4-nitrophenyl-63-α-D-maltotriosyl-maltotriose (BPNPG3G3), coupled with the ancillary enzymes α-glucosidase and β-glucosidase. Upon hydrolysis of the substrate at the 1,6-α-linkage by pullulanase or limit-dextrinase, the released 4-nitrophenyl-β-maltotrioside is immediately hydrolysed to glucose and 4-nitrophenol by the concerted action of the α-glucosidase and β-glucosidase enzymes in the reagent mixture. The reaction is terminated and phenolate ions are developed by addition of dilute alkali. The absorbance is read at 400 nm and the value obtained correlates directly with pullulanase activity.
All reagents stable for > 1 year after preparation
Very specific
Simple format
Standard included
Other Products
ProteoSpin™ Inclusion Body Protein Isolation Kits
Product Info
Document
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.
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
Propargyl-PEG9-alcohol is a crosslinker that can participate in copper catalyzed azide-alkyne Click Chemistry reactions to form stable triazole linkage. The PEG spacer increases the hydrophilicity of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
Propargyl-PEG9-alcohol is a crosslinker that can participate in copper catalyzed azide-alkyne Click Chemistry reactions to form stable triazole linkage. The PEG spacer increases the hydrophilicity of the molecule in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
2-(Propargyl-PEG4-amido)-1,3bis(PEG1-methyl ester) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The methyl ester groups can be hydrolyzed, reduced, or substituted under different conditions.
Document
2-(Propargyl-PEG4-amido)-1,3bis(PEG1-methyl ester) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The methyl ester groups can be hydrolyzed, reduced, or substituted under different conditions.
