Description
Specifications
| Clone | IHC409 |
| Source | Mouse Monoclonal |
| Positive Control | Colon, Colon Carcinoma |
| Dilution Range | 1:200 |
MutL Homolog 1 (MLH1) is a protein involved in the mismatch-repair pathway. This protein is commonly associated with hereditary non-polyposis colorectal cancer, as the MLH1 gene is frequently mutated in patients with this cancer. Studies have shown MLH1 to be deficient in a high percentage of patients with microsatellite instability, as well as endometrial and ovarian cancers. Use of Anti-MLH1 is optimized when paired in an IHC panel with MSH6, MSH2, and PMS2. Anti-MLH1 is useful in the detection of MLH1 in a number of normal and neoplastic tissues, and for identifying a loss of MLH1 in tumors that are microsatellite-unstable.
| Clone | IHC409 |
| Source | Mouse Monoclonal |
| Positive Control | Colon, Colon Carcinoma |
| Dilution Range | 1:200 |
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.
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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 |
The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.
The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic approach (for example, BioDynami DNA fragmentation enzymes, Cat.# 40061 and Cat.# 40062) and mechanical approaches such as sonication and nebulization etc.
NGS Low Input DNA Library Prep Kit Workflow
Three index types are available for the NGS Low Input DNA Library Prep Kit of illumina platform:
Non-index (Cat.# 30022): Libraries do not have index.
Index (Cat.# 30024): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify the low input DNA libraries. Library multiplexing up to 48 samples is possible. Index information can be downloaded here.
Unique dual index (Cat.# 30025): Library multiplexing up to 96 low input DNA libraries is possible with unique dual indexes with our Four-Base Difference Index System. The system allows us to make indexes for the libraries that have at least 4 bases different from each other in the 8-base barcode length. Our unique dual index primers can reduce sequencing errors such as de-multiplexing errors, amplification errors, mis-assignment of reads, index cross-contamination, and also index hopping. The kit includes 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34024).
Kit advantages:
Comparison of library conversion efficiency under the same NGS library preparation condition. Input DNA amounts are 1 ng and 10 ng, respectively. DNA was mechanical sheared with Covaris before library prep. BioDynami kit: NGS Low Input DNA Library Prep Kit (Cat. #30022).
Comparison of library yield under the same NGS library prep condition. Input DNA amounts are 1 ng, 10 ng, and 100 ng. DNA was mechanical sheared with Covaris before library prep. BioDynami kit (Cat. #30022). PCR cycle numbers were indicated.
