Suspend 111 g in 1 L of distilled or deionized water. Heat with frequent agitation and boil to completely dissolve the powder. Distribute into flasks. Autoclave at 121°C for 15 minutes. Cool to 50°C and pour into sterile Petri dishes.
Formula (per liter)
Pancreatic digest of casein—————5g Pancreatic digest of animal tissue——5g Beef extract————————————1g D-mannitol————————————-10g Sodium chloride—————————–75g Agar———————————————15g Phenolred——————————–0.025g Final pH———————————–7.4±0.2
Storage
Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
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
The Kit integrates phenol/guanidine-based lysis and silica membrane purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of samples and inhibit RNases. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable extracting up to 200μg total purified RNA from less than 100mg animal, plant, fungal samples,bacteria, cells and other samples.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA (include miRNA) from 100mg lipid tissue, tissue, cell, plant, body fluids using columns and MagZol reagent
Applications
RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Tissue samples (10-100mg) are homogenized in MagZol Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30-100µl of RNase-free water.
Advantages
Efficient DNA removal – one step RNA extraction can effectively remove genomic DNA
High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
Fast – several samples can be extracted in 30 minutes
High applicability – samples including animals, plants, bacteria, cells, etc.
High output – enable to process large samples and the yield can be up to 200μg
Kit Contents
Contents
R413002
D413003
Purification Times
50 Preps
250 Preps
HiPure RNA Mini Columns
50
250
2ml Collection Tubes
50
250
MagZol Reagent
60 ml
270 ml
Buffer RW1
50 ml
200 ml
Buffer RW2*
20 ml
50 ml
RNase Free Water
10 ml
30 ml
Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Experiment Data
Document
The Kit integrates phenol/guanidine-based lysis and silica membrane purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of samples and inhibit RNases. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable extracting up to 200μg total purified RNA from less than 100mg animal, plant, fungal samples,bacteria, cells and other samples.
t-Boc-aminooxy-PEG4-propargyl is a click chemistry tool containing a propargyl group and t-Boc-aminooxy group. Propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. T-Boc-aminooxy can be deprotected under mild acidic conditions and then can react with an aldehyde or ketone group to form a linkage. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
t-Boc-aminooxy-PEG4-propargyl is a click chemistry tool containing a propargyl group and t-Boc-aminooxy group. Propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. T-Boc-aminooxy can be deprotected under mild acidic conditions and then can react with an aldehyde or ketone group to form a linkage. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
