This monoclonal mouse antibody to human pancreatic secretory trypsin inhibitor domain (hPSTI) recognizes native hPSTI (soluble as well as hPSTI in the pSKAN phagemid pIII-hPSTI fusion protein).
The anti-hPSTI antibody was specifically developed for our pSKAN Phagemid Display System, but can also be useful in other applications related to the human pancreatic secretory trypsin inhibitor (hPSTI).
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PAGE of pSKAN phages presenting different hPSTI-pIII fusion proteins followed by Western blot with anti-pIII (lane 1-5); M13K07 wild type phage as control (lane M13). Two bands are visible with pSKAN phages: one for pIII and one for the larger hybrid protein hPSTI-pIII. In M13 wild type phages only one band is detected resulting from pIII.
The T7 RNA polymerase (T7 RNAP) expression system originates from the bacteriophage DNA-dependent RNA polymerase. In 1985 the first described T7 RNAP expression system was developed for Escherichia coli (Tabour and Richardson, 1985). Advantages of this system are the stringent selectivity and the high transcriptional activity so that it is possible to lead to a saturation of the protein-synthesizing machinery in E. coli. Consequently 50% or more of the total cellular protein can consist of the desired protein (Studier and Moffatt, 1986).
The T7 RNAP expression system for Bacillus megaterium combines the features of this system in E. coli with the above mentioned regulation by the xylose operon. This system is based on two parallel-replicating plasmids: pT7-RNAP and pPT7 (Gamer et al. 2009).
In addition to the t7 rnap gene under control of the strong xylA promoter pT7-RNAP contains the genes of ampicillin and chloramphenicol for easy selection in E. coli (AmpR) and B. megaterium (CmR).
pPT7 is responsible for the T7 RNAP-dependent expression of the target gene. Downstream of the T7 promoter it comprises a multiple cloning site with ten unique restriction enzyme cleaving sites. Additionally the plasmid comprises two resistances to ampicillin (in E.coli) and tetracycline (in B. megaterium).
For your convenience MoBiTec offers B. megaterium protoplasts pretransformed with pT7-RNAP, so that you just have to insert your gene of interest into pPT7 and transform the pretransformed protoplasts with this plasmid.
For control purposes the GFP-expressing vector pPT7-GFP is included in the kit.
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The Dri-Capsules are used in conjunction with Isohelix SK-1 Buccal Swabs to stabilize the swabs for long term storage of DNA samples. After the buccal DNA samples is taken, a single Dri-Capsule is placed in the collection tube, above the swab (a portion of the swab shaft separates the capsule from the head of the swab).
The Dri-Capsules are ideal when samples are being taken at remote locations as no refrigeration or freezing is required and use of the Dri-Capsules does not require trained personnel.
A kit containing both the buccal swabs and the Dri-Capsules is available – click here for details
Bacillus subtilis is an attractive host for the production of recombinant proteins. There are several reasons for this: it is capable of secreting functional extracellular proteins directly into the culture medium (at present, about 60% of the commercially available enzymes are produced by Bacillus species); it has no significant bias in codon usage; and a large body of information concerning transcription, translation, protein folding and secretion mechanisms, genetic manipulation and large-scale fermentation has been acquired.
The Bacillus subtilis expression vectors are plasmid based expression vectors for highly efficient intra- and extracellular production of recombinant proteins in Bacillus subtilis. The pHT vectors use the strong promoter preceding the groESL operon of Bacillus subtilis fused to the lac operator allowing their induction by addition of IPTG. The pAL Vectors contain the cold inducible des promoter of Bacillus subtilis. pAL10 and pAL12 allow for intracellular synthesis of recombinant proteins without the need for a cost intensive inductor.
We are also pleased to offer Bacillus subtilis host strains for intracellular expression as well as for secretion vectors.
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Magnetofection is our term for Magnetic Transfection. Magnetofection associates nucleic acids or other vectors with magnetic nanoparticles coated with cationic molecules. The magnetic nanoparticles are complexed with vectors (DNA, siRNA, oligos, virus, etc.) by salt-induced colloidal aggregation and electrostatic interaction. The resulting molecular complexes are then attracted to target cells by an appropriate magnetic field (Magnetofection Magnet), and enter the cells by endocytosis. In this manner, the complete applied vector dose gets concentrated on the cells within a few minutes. As a result, 100% of the cells come into contact with a significant vector dose.
Special Magnetofection reagents have been developed for many applications. These include PolyMag (for general use); CombiMag (for use in combination with standard lipid based transfection reagents); ViroMag and ViroMag R/L (to increase transduction efficiency of viral vectors); NeuroMag (specialized for use with neurons); and SilenceMag (siRNA delivery).
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When isolating DNA from buccal swabs, it is important to use a DNA isolation kit that has been optimized for use with buccal swabs. The Isohelix Buccal DNA Isolation Kit is one example of a kit that has been optimized for use with buccal DNA swabs. It has been specifically formulated to produce high DNA yield and purity from buccal swabs. The kits have been fully optimised for use on buccal cell samples and offer reduced handling times, increased DNA yields and many other important technical benefits for their use in manual, 96-well or other high throughput formats. Furthermore, various formats are available including the DNA isolation kit by itself, or combined with one of two formats of Isohelix Buccal Swabs: SK-1S (packaged with a 5ml collection tube) or SK-2S (packaged with a 2ml collection tube and special cap to facilitate removal of the swab head).
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Taq polymerase is a heat stable DNA polymerase derived from Thermus aquaticus. It is extensively used in the polymerase chain reaction or PCR. The DNA products formed with Taq polymerase have an adenine (A) overhang which is advantageous for TA cloning.
DFS (DNA Free Sensitive) Taq DNA Polymerase is a thermostable DNA Polymerase of approximately 94 kDa isolated from Thermus aquaticus strain YT-1. This unmodified Taq replicates DNA at 72°C. The DFS-Taq Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5′–> 3′ direction in the presence of magnesium ions and possesses a 5′–> 3′ exonuclease activity. The DNA-free Taq Polymerase is highly purified and is free of nonspecific endo- or exonucleases.
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DFS (DNA Free Sensitive) Taq DNA Polymerase is highly purified and is free of nonspecific endo- or exonucleases. This Taq DNA Polymerase is absolutely free of bacterial DNA
DFS (DNA Free Sensitive) Taq DNA Polymerase is suitible for: Standard PCR of DNA fragments up to 5 kb; Realtime PCR; Primer extension reaction; PCR with bacterial DNA
FEATURES: High sensitivity, only 6 molecules template required; Efficient amplification of single copy genes; High product yields
1 – without adding E.coli DNA (Test)
2 – with adding E.coli DNA (control)
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Our HiMaX range of Electroporation cuvettes are designed to maximise electroporation efficiencies for Bacteria, Yeast, Insect, Plant and Mammalian cells. Our rigorous quality control procedures insure reproducible results every time.
Cap Design: The cap has been designed to improve aseptic handling techniques, while the lip and positive seal reduces potential aerosol and contamination issues; Low Dead Volumes: All 1mm and 2mm cuvettes have a tapered V bottom so that reduced sample volumes can be used while aiding sample pick up and minimising dead volumes; Compatibility: The cuvettes are compatible with most electroporation systems; Size Range and Colour Coded: Available in 1mm, 2mm and 4mm gap sizes with individual colour caps; Bio-Controlled: All batches are checked to optimise the Bio and Transfection compatibility, with stringent use of high quality grade polycarbonate and High grade chemicals to ensure consistent uniform pulse generation and improved gene transfer; Sterile Packaged: Every cuvette is guaranteed sterile, packed using gamma irradiation and has an simple tear wrapper for easy access when you need it; High Tolerance Moulding: The moulding process ensures extremely high tolerances so that the electrodes have a consistent gap and parallel configuration. The electrodes are also cleaned chemically and physically to fully optimise the cuvette for high transformation efficiencies.
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ViroMag is a viral gene delivery reagent based on Magnetofection technology.
Magnetofection is a new technology which uses magnetic force to concentrate viral vectors (which have been complexed with magnetic particles) on the cells within a few minutes so that 100% of cells are in contact with the viral vector. ViroMag thus significantly increases transduction efficiency and accelerates the transduction process.
Magnetofection and ViroMag can be used with numerous cell types including a variety of immortalized and primary cells.
ViroMag is designed to be used in conjunction with all viral vectors to:
Accelerate the transduction process; Improve virus infectivity with low vector doses; Increase transduction efficiency; Infect non-permissive cells; Concentrate the complete viral dose on the target cells very rapidly; Target transduction to a specific area (magnetic targeting); Synchronize cell adsorption/infection
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