MoBiTec releases new Cell Viability and
Cytotoxicity Assays brochure

The new brochure provides an overview on a wide range of cell viability and cytotoxicity kits. It comprises not only kits made by MoBiTec but also kits from manufactures distributed by MoBiTec in Germany, e.g. Molecular Probes, Amresco and AnaSpec.
The kits can be used to examine animal cells, bacteria, yeast, and fungi, and differ in their detection methods. All kits are clearly arranged and described so that you can easily find the best kit for your application.

Please download our brochure as PDF file.

Upgrade Your Real Time PCR for more Reliable Results

Is your Gene Expression underestimated due to residual RNA inhibiting the qPCR ?

Features of Tli RNaseH added premixes:

• Improved quantification by the addition of thermostable Tli RNaseH (No RNase pre-treatment needed)
  
• Quantification of various targets with broader dynamic range

SYBR^® Premix Ex Taq™ (Tli RNaseH Plus)

• Higher Extension Rates: allows faster reaction times
  
• Amplification of Longer Fragments: ability to amplify fragments larger than
  500 bp

SYBR^® Premix Ex Taq™ II (Tli RNaseH Plus)

• Higher Specificity: no primer dimers or non-specific amplification
  
• Excellent Cq values

All Takara qPCR Premixes contain high and low concentration ROX reference dye in separate tubes for use in any real time thermocycler.

EHEC Real Time PCR Kit, CE Marked

Intended Use
Enterohemorrhagic E. Coli (EHEC) real time PCR kit is used for the detection of Enterohemorrhagic E. Coli (EHEC) in excreta or water samples by using real time PCR systems.

Introduction
Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is the serotype most commonly associated with hemorrhagic colitis and the hemolytic uremic syndrome. It is phenotypically distinct from typical E. coli, in that they do not ferment sorbitol and do not exhibit glucuronidase (GUD) activity. They are also easily identified by their ability to produce Shiga toxins (Stx) and by the presence of the somatic O157 and the flagellar H7 antigens. As a result, these traits are extensively utilized in methods used to isolate and detect O157:H7 in foods. However, new variant strains of EHEC, particularly O157:H7, are being isolated more frequently from foods, animals and humans worldwide. These include sorbitol-fermenting variants, strains that exhibit GUD, non-motile, toxigenic variants; serological variants; toxin non-producing strains, etc. These variants carry most of the trait EHEC virulence markers, so are potentially pathogenic and some have already been implicated in illness; however, due to changes in trait genotypic and phenotypic markers, EHEC variants elude detection by current testing methods.

Cat# DD-0095-01-ZJ
（Ⅰ）LightCycler 1.0； (Internal Control is not included for this system)
（Ⅱ）LightCycler2.0，

Please download manual (PDF, 172 KB)

Cat# DD-0095-02-ZJ
（Ⅲ）PE5700，MJ-Opticon etc. single color systems；
（Ⅳ）ABI7000，ABI7300，ABI7500，ABI7900，ABI StepOne,StepOne plus, MJ
Opticon2，MJ-chromo4，MX3000P，MX3005P，Smart CyclerII，Rotor-Gene6000,LightCyclerR480；iCycler iQ4，iCycler iQ5, etc, multi-color systems.

Please download manual (PDF, 242 KB)

Release of MoBiTec Newsletter VIII

The new international Newsletter VIII includes:

• The new photoactivatable and photoconvertible fluorescent protein mIris FP, an advanced version of the well known EosFP.
  
• M-Beads Magnetic silica beads for diverse proteomic applications
  
• High yield CMV promoter-based constitutive expression vectors
  
• Anti-hPSTI and Anti-g3p (pIII) antibodies
  
• Highly efficient BactoLyse bacteria protein extraction reagent
  
• NeuroTrans and siRTrans transfection reagents for efficient transfection into primary neurons and transfection of siRNA into mammalian cells
  
• New plastic ware for microscopy: Imaging Dishes and ImmunoSelect^® Adhesion Slides
  
• Optimized high performance (hp) vectors for protein expression in Bacillus megaterium

Download the new newsletter as pdf (752 kb) or request a paper copy for free.

Photoactivatable and photoconvertible fluorescent protein - mIrisFP

IrisFP is a photoactivatable fluorescent protein that combines irreversible photoconversion from a green- to a red-emitting form with reversible photo-switching between a fluorescent and a non-fluorescent state in both forms. mIrisFP is a monomeric variant. mIrisFP multiple photo-activation modes can be used for pulse-chase experiments combined with subdiffraction-resolution imaging in living cells by using dual-color photo-activation localization microscopy (PALM).

Monomeric mIrisFP has excellent properties as a genetically encoded fluorescence marker. mIrisFP fluorescence can be reversibly switched on and off (useful for high-resolution microscopy, PALM), but also allows for the non-reversible photo-conversion from green to red (useful for dynamic measurements in living cells). The dual photo-activation capability of mIrisFP offers enhanced flexibility and also enables the combination of pulse-chase experiments with super-resolution imaging.

Thermostable, photoconvertible fluorescent protein – mEosFP(Thermostab)

mEosFP(Thermostab) is an advanced variant of the green to red photoconvertible fluorescent protein EosFP. The marker was optimized for functional expression at 37 °C, especially in fusion with other proteins. mEosFP(Thermostab) retains the monomeric nature of its predecessor and shows an excellent performance in fusion even with demanding partners such as tubulin. mEosFP(Thermostab) can be efficiently converted from green to red by a light pulse at wavelengths between 360 and 430 nm.

For more information please visit our photoconvertible fluorescent proteins.

Superior accessories for fluorescence microscopy and imaging

Our cell culture and microscopy ware for imaging applications provides a great opportunity, when living or fixed cells are in the focus of your research. In addition to our ImmunoSelect® Adhesion slides we are now offering plastic ware for fluorescence microscopy. The range comprises Imaging Dishes with and without µgrids and diverse Imaging Plates with different surfaces and made out of variable materials. Most products provide Tissue Culture surfaces and promise high image quality, cell adhesion and differentiation.

Until March 31 you can get a free sample of Imaging Dish CG or Adhesion slides on request. Please contact us or your local distributor.

Our whole range of fluorescence microscopy accessories can be found here.

Mobicol brochure online available

Mobicols are practical plastic tools suitable for diverse lab applications. In our new brochure you can get an overview on miscellaneous Mobicols and their applications. Different chapters describe the plastic columns and their usage:

Chapter I       Mobicol – a practical tool for every lab
Chapter II      Spin columns for your own matrices or other compounds
Chapter III     Matrix-filled MobiSpin columns for purification of nucleic acids
Chapter IV    Mobicols for affinity purification of biomolecules
Chapter V     Mobicols with enzymatically active matrices

Please download the new Mobicol brochure as pdf.

With the new TransIT-PROTM Transfection Kit critical Stages are achieved faste

Mirus Bio now offers a new transfection kit ideal for biotherapeutic protein production in large pharma and biotech settings. Mirus Bio’s new TransIT-PRO™ Transfection Kit will decrease time to produce usable protein by maximizing target protein yields through transient transfection. TransIT-PRO™ Transfection Kit uses animal origin free components designed for high and reproducible protein yield in suspension CHO and 293 derived cells. Since it is compatible with varied media formulations, the same media can be used for both transient and stable expression. TransIT-PRO™ outperforms linear PEI in protein yield and reproducibility while providing a cost-effective alternative to FreeStyle™ MAX.

Features

• Achieve high protein yield in suspension CHO and 293 cells
• Experience compatibility with multiple CHO media formulations
• Obtain reproducible protein expression with minimal optimization

For more information please visit our Cell Transfection Reagents.

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SensoLyte^® Biotin Quantitation Assay Kit – Fluorimetric Biotin Quantification Assay

AnaSpec is pleased to announce the release of the FRET-based biotin quanti kit. The SensoLyte® Biotin Quantitation Assay Kit provides a convenient way to estimate the efficiency of biotinylation for biotin-protein conjugates. The kit also can be used to quantitate biotin concentration in a solution. In the first step, Reagent A, which is tagged with a quencher dye, is briefly incubated with biotin samples/standards. In the second step, fluorescent biotin, Reagent B, is added to assay mixture. The assay can detect between 2 to 20 picomoles of biotin in a sample (Fig. 1). A biotinylated IgG for use as a positive control and a protease for optional protein digestion to expose hidden biotin groups are also included in the kit.

Figure 1. Biocytin standard calibration curve. Serial dilutions of Biocytin were mixed with Reagent A, followed by Reagent B. After 5 minute incubation, fluorescence was measured at Ex/Em=490/520 nm  (FlexStation 384II).

Biotin and avidin (or streptavidin) bind non-covalently with a higher binding affinity than most antigen-antibody interactions.^{1, 2} This very tight binding makes labeling proteins with biotin a useful tool for applications such as affinity chromatography and immunoanalytical methods. Reaction conditions for biotinylation are chosen such that the target molecule (e.g. an antibody) is labeled with sufficient biotin residues to purify or detect the molecule, but not too much that the biotin will interfere with the function of the molecule.

Cys-containing β-Amyloid

The hallmark of Alzheimer’s disease (AD) pathology includes β-sheet aggregates of β-amyloid peptides in senile plaques and hyperphosphorylated tau protein in neurofibrillary tangles (NFT).^1-2 Recent publications have reported the use of Cys-containing mutants as models for aggregation studies.^3-5 Shivaprasad and Wetzel employ “the use of disulfide bond cross-linking to probe the fold within the core and the packing interactions between beta-sheets.” Among the Cys mutants they studied is the S26C (Ser substituted with Cys at position 26).³ Upon oxidation of the S26C monomer, cysteines are capable of forming intermolecular disulfide bond creating the S26C dimer.^3-4 Using this synthetic dimer, Hu, et al. show that it behaves similarly to β-amyloid dimer-containing human CSF, suggesting that amyloid β dimers may be the earliest synaptic disrupting species in AD.⁴
As the supplier of the world’s largest collection of β-amyloid GO™ (catalog) Peptides, AnaSpec is pleased to add to this list, β-amyloid (1-40) S26C; β-amyloid (1-40) S26C dimer; β-amyloid (1-42) S26C and other Cys-containing β-amyloid peptides (Cys on the N or C-terminus as well as in the internal sequence).
Besides using these Cys-containing β-amyloid peptides for dimerization, the availability of the cysteine’s thiol group also allows researchers the flexibility of reacting these peptides with any maleimide containing fluorescent dyes or biomolecules. For example, the thiol group of Cys-β-amyloid (1-40) can react with HiLyte Fluor™ 488, C2 maleimide to form HiLyte Fluor™ 488-Cys-β-amyloid (1-40) [Cys(HiLyte Fluor™ 488)- DAEFRHDSGYEVHHQKL VFFAEDVGSNKGAIIGLMVGGVV].

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References

Quantitation Assays:

1. Angerer, L. et al. Cell. 9, 81-90 (1976).
   
2. Gitlin,G. et al. Biochem. J. 242,923 (1987).

β-Amyloid peptides:

1. Khachaturian, ZS. Arch. Neurol. 42, 1097 (1985).
   
2. Mirra, SS. et al. Neurol. 41, 479 (1991).
   
3. Shivaprasad, S. and R. Wetzel. J. Biol. Chem. 281, 993 (2006).
   
4. Hu, N-W. et al. Brain doi:10.1093/brain/awn174.
   
5. Shankar, GM. et al. Nat. Med. 14, 837 (2008).

Click-Chemistry

Click-chemistry was first introduced by K. Berry Sharpless in 2001. It describes a chemical reaction which builds substances by joining small units together. Click-reactions are fast and purposeful with high yields.
The most popular click-reaction is azide alkyne Huisgen cycloaddition with Copper (Cu) as catalyst at room temperature.
Now you can use this highly efficient technology for labeling of DNA or RNA via solid-phase synthesis or PCR. Furthermore, it is possible to introduce up to three different modifications onto one position of the same DNA strand. Of course, we offer different kinds of non-fluorescent and fluorescent labels.

For more information please visit our Click-Chemistry overview.

SapphireAmp^® Fast PCR Mix

Features:

• Faster than standard Taq: reactions completed in1/2 the time of conventional Taq
  
• Restriction enzyme digestion can be directly performed on PCR products in the PCR buffer
  
• PCR reaction can be directly loaded onto a gel without purification or addition of other reagents
  
• Save time and eliminate purification steps with no reduction in yield

Description:

Fig.: Template: Human Genome 100ng/50mL Targets: 1: p53 0.5 kb (54%); 2: FFAR2 1.0 kb (59%); 3: DCLRE1A 2.0 kb (38%); 4: p53 4.2 kb (48%); 5: IGF2R 5.9 kb (50%)

SapphireAmp^® Fast PCR Master Mix contains a hot start PCR enzyme, optimized buffer, dNTP mixture, gel loading dye (blue) and a density reagent in a 2X premix. PCR reactions with this mix can be completed in half the time required for conventional Taq amplifications. Amplification of human genomic DNA targets of 2 kb in length can be completed in 1 hour, and amplifications of human genomic DNA targets of up to 6 kb are possible using Sapphire-Amp™. This product is well-suited for E. coli-based colony PCR to check for inserts of up to ~5 kb, and colony checks can be completed in about 1 hour. SapphireAmp^® Fast PCR Master Mix reactions can be assembled by simply adding primer and DNA template to the premix. Reactions performed with this mix can be loaded directly onto a gel for electrophoresis, and restriction digestion of PCR products is possible in SapphireAmp™ reaction buffer. SapphireAmp^® Fast PCR Master Mix offers a powerful, fast, flexible and convenient alternative for both simple and complex PCR applications.

Table: SapphireAmp^® Master Mix saves time.

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New Vector Systems Brochure available from MoBiTec

Our new brochure focuses on gene expression in different microorganisms like Escherichia coli, Bacillus megaterium, Bacillus subtilis, Lactococcus lactis and yeast. Furthermore, it includes expression systems for mammalian cells as well as vector systems for many other purposes.

Content:

• NICE^® Expression System for Lactococcus lactis
  
• Bacillus megaterium Expression Systems
  
• Bacillus subtilis Expression Vectors
  
• pBacTag Tagging Vectors
  
• Yeast Expression Vectors
  
• pORF-CLONE Vector System
  
• pPICHOLI Shuttle Vector System
  
• Mammalian Expression Vectors
  
• CMV Expression Vectors
  
• Fusion Protein Cloning Systems
  
• BRP Plasmids and Competent BRP Cells
  
• PCR Cloning Vector p3T
  
• Poly(His)-tag Cloning Vector pEG-His1
  
• Exontrap Cloning Vector
  
• ssDNA-Production and Expression in pMEX
  
• Promoter-Trap Vector pBBR RESO
  
• Broad-Host-Range Vectors pBBR122 and pBHR1
  
• Multiple Cloning Site Vector pMCS5
  
• Standard Cloning Vectors
  
• EosFP – Green to Red Photoconvertible Fluorescent Protein
  
• Related Products

Please download the new Vector Systems Brochure (PDF, 3879 KB) !

AMRESCO’s InvestiGator including products for:

• Protein Electrophoresis
  
• Western Blotting
  
• Proteomics

Download the new Newsletter as PDF (1600KB) or request a paper copy for free.

High Yield Gene Expression with the new T7 RNA Polymerase Gene Expression System!

This system combines the benefits of the tightly regulated and strong T7 RNA polymerase gene expression system and alkaline protease free Bacillus megaterium.

The T7 RNAP gene expression system is tightly regulated and efficiently inducible through the xylA operon and the strong T7 RNAP promoter resulting in stable, high yield protein production up to 8-fold increased in comparison to common xylose inducible expression (Fig. 1).

The system is based on two parallel-replicating plasmids: pT7-RNAP and pP_T7.
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 (Amp^R) and B. megaterium (Cm^R).
pP_T7 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 cleavage sites. Additionally, the plasmid carries two resistances to ampicillin (in E. coli) and tetracycline (in B. megaterium).

Fig. 1 Comparison of GFP amounts produced by variant expression systems. GFP produced by B. megaterium carrying the common xylose-regulated Plasmide pRBBm34 (square), B. megaterium transformed with the T7 expression system plasmids (circle) and B.megaterium with the T7 expression system plasmids treated with rifampicin. (Gamer et al., 2009)

Pretransformed B. megaterium protoplasts!

For your convenience we offer B. megaterium protoplasts pretransformed with pT7-RNAP, so that you just have to insert your gene of interest into pP_T7 and transform the pretransformed protoplasts with this plasmid.

Absolute Detection of Apoptotic Transformation
- Violet Ratiometric Membrane Asymmetry Probe/Dead Cell Apoptosis Kit

What it is

What it offers

• Robust assay - ratiometric probe is independent of probe concentration, cell size, and instrument variation
  
• Easy to use - 5-minute incubation, with no wash required
  
• Flexible experimental design - easily combined with other testing reagents
  

How it works

This assay employs a novel dye (F2N12S) that incorporates selectively into the outer leaflet of the plasma membrane. The dye exhibits dual fluorescence emission bands corresponding to 530 nm and 585 nm, producing a ratiometric response to variations in plasma membrane surface charge during early apoptosis that is detected by flow cytometry. The kit also includes SYTOX^® AADvanced™ Dead Cell Stain for discrimination of dead cells from live cells. Unlike annexin V conjugates, this assay requires no special buffers or wash steps and is less affected by cell membrane damage commonly associated with the removal of adherent cells, thereby improving data quality.

DNAgard™ for DNA stabilization in tissues and cells

Biomatrica introduces the DNAgard™ a new product for DNA stabilization in tissues and cells. DNAgard™ stabilizes genomic DNA in cultured cells or animal tissues for at least 2 months in liquid storage format at room temperature. For longer term storage dry-down is required.

DNAgard™ is designed for immediate stabilization of DNA from intact cells and tissues with the convenience of room temperature shipping, processing and storage. The liquid storage reagent rapidly permeates cell membranes to stabilize and protect genomic DNA.

With its excellent properties DNAgard™ is ideal for field collection and tissue harvesting. Of course DNA protected using DNAgard™ is suitable for use in many downstream applications, including long-range PCR and Real Time PCR.

For longer storage, samples can be dried down and stored at room temperature for at least 1 year* (*accelerated aging). Dry samples can be easily recovered by simple rehydration and are ready for DNA extraction

K-2500s PIP3 Mass ELISA Kit

Echelon Biosciences introduces the K-2500s PIP3 Mass ELISA. This kit is 10x more sensitive to PIP3 than our K-2500 assay and offers less cross reactivity with PIP's (most specifically PI(3,4)P2).

K-2500s - PIP3 Mass ELISA

The K-2500s uses different PIP3 Detection protein and buffers than the
K-2500 PIP3 Mass Elisa Kit, resulting in 10x more sensitivity and selectivity to PIP3.

Class I Phosphoinositide 3-kinases (PI3-Kinase, PI3-K), are ubiquitously expressed lipid kinases which phosphorylate PtdIns(4,5)P2 at the 3'-hydroxyl of the inositol ring producing PtdIns(3,4,5)P3 (PIP3). PIP3 has shown to be an important lipid second messenger in multiple cell signalling pathways such as proliferation, metabolism, calcium release, and survival. Due to these interesting biological roles, PI3-K and PIP3 have become attractive targets for research and drug development.

The PIP3 Mass ELISA Kit allows the using to determine PI3-K activity by quantifying the amount of PIP3 found in cells. After following a non-radioactive lipid extraction protocol, the PIP3 samples obtained from millions of cells are detected on a competitive ELISA. The cellular PIP3 samples are mixed and incubated with a PIP3 Detector protein, then added to a PIP3-coated microplate for competitive binding. A peroxidase-linked secondary detector and colorimetric detection is used to detect the PIP3 protein binding to the plate. The colorimetric signal is inversely proportional to the amount of PIP3 obtained.