Supplier Focus: SignalChem’s Active Kinase Mutants

Phosphorylation is a post translational modification (PTM) of proteins and lipids and it represents one of the most efficient ways to regulate protein and lipid function in cells. Protein kinases are one of the largest classes of enzymes and they are responsible for phosphorylating proteins. Kinases themselves are turned on or off by growth factors, cytokines and hormones to regulate a diverse set of downstream cell signaling pathways.

Defective cell signaling is implicated in several inherited disorders and diseases, such as cancer, diabetes, and cardiovascular disease. Mutations in kinases can lead to defective signaling and indeed, there are many characterized kinase mutants that play a causal role in disease. Moreover, cancers can acquire drug-resistance mutations such as in the case of EGFR (T790M), which can diminish the efficacy of conventional and targeted therapies.

SignalChem strives to provide researchers with the right tools for their drug discovery efforts and these include disease-related kinase mutants. They are leading the way by providing one of the largest libraries of Active Kinase Mutant enzymes in the industry. They are continually developing new clinically relevant kinase mutants.

For a list of all kinase from SignalChem check out the following link.


Where can I find more information about SignalChem?

Visit the manufacturer page at www.stratech.co.uk/signalchem, email info@stratech.co.uk or call +44 (0) 1638 782600

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.

FIRST SCOTTISH CYTOMETRY WORKSHOP

Boyd Orr Building, University Avenue, University of Glasgow, Glasgow G12 8QQ
June 24th-25th 2015

ISAC’s Live Education Task Force, in conjunction with flowcytometryUK and the Royal
Microscopical Society, will run a two-day hands-on flow cytometry course on Wednesday
24th and Thursday 25th June immediately prior to this year’s CYTO Conference.

CYTOFlyer

This course is aimed at those relatively new to cytometry or to those who wish to broaden
their knowledge. There will be some introductory talks and then seven wet-labs of which
delegates can choose four to attend. Each wet-lab session will be three hours. The wet labs will be open to 50 registered particpants on a first come, first served basis.

WORKSHOP MODULES

  1. DNA content and cell cycle analysis (Mammalian and plant)
  2. Cell Proliferation (Nucleotide uptake and dye dilution)
  3. Multicolour phenotyping and compensation
  4. Imaging flow cytometry (Principles and applications)
  5. Cell Viability and cell death including apoptosis
  6. Functional cytometry (Titration, receptor quantification)
  7. Electrostatic cell sorting

Registration and further information:
http://www.rms.org.uk/events/Forthcoming_Events/practical-flow-cytometry-course
Registration fees: Academic £200, Non-academic £300

NEW Product: AAVanced™ Concentration Reagent – One-step rAAV particle isolationsys

Easiest method for rAAV isolation

  • Easy to use, single reagent
  • Isolate rAAV directly from media
  • No cell lysates or centrifugation
  • Saves time and cost-effective
  • Compatible with all serotypes

AAVanced™ Concentration Reagent

One-step rAAV particle isolation from packaging cell media
Traditional rAAV production methods requires multiple steps, including cell lysis and CsCl2 ultra-high-speed density gradient centrifugation, chromatography, or binding to affinity matrix columns. These are difficult to setup, time-consuming, and require specialized equipment for isolation of high-purity rAAV for in vivo experiments. As many studies have shown that different rAAV serotypes are efficiently secreted into the culture medium of transfected 293T cells during rAAV packaging, SBI has developed a novel reagent to facilitate fast, high-quality rAAV isolation.

   

Easiest Protocol to Isolate rAAV Particles

SBI has developed an innovative solution called AAVanced Concentration Reagent – a simple, one step rAAV concentration reagent for the isolation of rAAV particles from media. AAVanced Concentration Reagent is based on a proven nanoparticle technology and is specifically optimized for the precipitation of most AAV serotypes from culture medium. The rAAV virus produced using AAVanced Concentration Reagent has been tested successfully for in vitro and in vivo applications for multiple serotypes of AAV with no observed cytotoxic effects, which provides direct evidence for the utility of this reagent for demanding applications using rAAV. SBI’s AAVanced Concentration Reagent significantly reduces the complexity and time required for rAAV virus production, allowing researchers to focus on their research experiments rather than virus production.

AAVanced Concentration Reagent Produces Robust Virions

How does AAVance virus preps compare to tranditonal cell lysate preps?
To test this comparison, recombinant rAAV was packaged with a PGK-GFP expression shuttle vector. The rAAV particles were isolated either by the traditional cell lysis freeze-thaw with centifugation method or with the AAVanced Concentration Reagent from the packaging cell media. Equal amounts of rAAV were then added to HEK293 cells to test for infectivity based on GFP expression. Representative images of GFP expressing cells 10 days after infection.

AAVanced virus produces infectious virus


AAVanced Concentration Reagent

AAVanced Concentration Reagent (100 ml) 100 ml AAV100A-1
AAVanced Concentration Reagent (250 ml) 250 ml AAV110A-1

Online Resources


Where can I find more information about SBI?

Visit the manufacturer page at www.stratech.co.uk/SBI, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


NEW Product: ViaCheck™ 25% Viability Control

Stratech Scientific are pleased to announce an addition to the Bangs Laboratories line of ViaCheck products for QC of automated viability analyzers such as the Vi-CELL XR.  The new ViaCheck 25% Viability Control.

Inventory will be available soon please contact info@stratech.co.uk for further information.

Catalog Code Description Volume (mL) Product Data Sheet Material Safety Data Sheet
VC25B ViaCheck™ 25% Viability Control 20 PDS 706 MSDS VC Viability Control

Comparison Study of the Vi-CELL® XR Cell Viability Analyzer and Z2™ Coulter Counter  K.Kilbride, M. Meza, R. Anglea, CYTO 2014, FT. Lauderdale, FL, May 17-21, 2014


Where can I find more information about Bangs Laboratories?

Visit the manufacturer page at www.stratech.co.uk/bangs_laboratories, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


Product Focus: Z-TECT™ High Sensitivity (HS) In Vitro Research Kits

All Z-TECT™ HS In Vitro Research Kits were designed for Research-Use-Only (RUO) to provide maximum ease of use and efficiency for the end-user by offering a turnkey solution. Z-TECT™ HS Secondary Research Kits contain all the reagents and materials necessary for performing a direct or sandwich ELISA assay (microwell plates, adhesive plate sealers, buffers and detection reagents) and are compatible with standard lab research equipment (pipettes, incubator, microwell plate washer and reader).

Akrivis Z-TECT™ High Sensitivity (HS) In Vitro Research Kits

Catalogue No.

Product Name

Product Data Sheet

Brochure

TSK480-21112-H001 Detection Kit, Goat anti-Mouse, Z-TECT™ (HRP) PDF PDF
TSK480-21113-H001 Detection Kit, Goat anti-Rabbit, Z-TECT™ (HRP) PDF PDF
TSK480-24001-H001 Detection Kit, Streptavidin anti-Biotin, Z-TECT™ (HRP) PDF PDF

Where can I find more information about US Biological?

Visit the manufacturer page at www.stratech.co.uk/usbio, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


Product Focus: Biotium CF Dyes™ Range

CF™ dyes are a series of highly water-soluble fluorescent dyes spanning the visible and near-infrared (NIR) spectrum for labeling biomolecules, especially proteins and nucleic acids. Developed by scientists at Biotium using new breakthrough chemistries, CF™ dyes rival or exceed the quality of other commercial dyes, such as Alexa Fluor® dyes, due to the following novel features.


New rhodamine chemistry

Rhodamine dyes are known for their excellent photostability and good fluorescence quantum yield; consequently several of the Alexa Fluor® dyes bear the rhodamine core structure.

Unfortunately, traditional rhodamine chemistry makes it difficult to extend the fluorescence wavelength to the far-red region and even more challenging in the near-IR region, especially for water-soluble dyes for bioconjugation. Recently, Biotium scientists discovered a new way to prepare novel rhodamine dyes of any fluorescence color from green to near-IR.

The new chemistry is a key element in the development of many of our CF™ dyes, particularly our far-red CF™ dyes, which are not only bright and water-soluble but also extremely photostable.


Excellent labeling efficiency

Reactive dyes for bioconjugation are generally susceptible to hydrolysis, which can cause problems for shipping, handling and storage and result in lower labeling efficiency. Heavily sulfonated dyes, such as the Alexa Fluor® dyes, DyLight® dyes and IRDye® are particularly hygroscopic, worsening the hydrolysis problem.

For example, the percent of active Alexa Fluor® 488 succinimidyl ester (SE) could be well below 50% by the time of application (Alexa Fluor 488 microscale labeling kit product information sheet, Invitrogen). In contrast, all of Biotium’s amine-reactive CF™ dyes have a relatively stable form of SE, which is more resistant to hydrolysis than the SE in many of the Alexa Fluor dyes. Accordingly, CF™ dye SE products generally give consistently higher labeling efficiency, thus providing users a better value.


Multi-color flexibility

Biotium currently offers 20 CF™ dyes spanning the visible, far-red, and near-infrared spectra, with additional colors in development. The CF™ dye product line includes reactive dyes, secondary antibodies, streptavidin, phallodin and other conjugates, and antibody labeling kits.


Unrivaled near-IR dyes

Near-IR dyes are typically much larger in size than dyes in the visible range. The large size often results in serious problems of low dye solubility, dye aggregation and poor fluorescence quantum yield. To overcome the problems, many commercial near-IR dyes, such as the near-IR Alexa Fluor® dyes, DyLight® dyes and IRDyes®, are prepared by placing a number of negatively charged sulfonate group on the dyes. While sulfonation improves dye solubility and fluorescence quantum yield to some degree, it creates another even more serious problem: non-specific binding of the bioconjugates prepared from the dyes.

For example, conjugation to a highly negatively charged dye can dramatically alter an antibody’s isoelectric point (iP), which is essential for maintaining specific antibody-antigen interaction. With this insight, Biotium scientists devised a revolutionary new approach to near-IR dye design that results in superior physical properties of the dyes without introducing an excessive amount of negative charge.

Biotium’s near-IR CF™ dyes are based on the core structure of either cyanine dyes or rhodamine dyes. Those core structures are modified such that the intramolecular mobility of the dyes is restricted, which leads to higher quantum yield and better water solubility without adding excessive charge. As a result, near-IR CF™ dyes are much brighter and more photostable than any other near-IR dyes. Most importantly, antibodies labeled with near-IR CF dyes™ give far better signal-to-noise ratio in immunostaining compared with antibody conjugates prepared with other commercial near-IR dyes.


Where can I find more information about US Biological?

Visit the manufacturer page at www.stratech.co.uk/usbio, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


Product Focus: XPack™ Technology – Engineer Exosome Protein Cargo for Delivery and Tracking

Exosome Protein Loading

  • Pack reporter proteins to track cargo
  • Make biotherapeutic exosomes
  • Generate bioluminescent exosomes
  • Create stable cell lines loading desired protein cargo

Exosomes are secreted nanoshuttles that facilitate communication between cells and organs and are found in various biofluids including blood, urine, amniotic fluid, breast milk, malignant ascites fluid, and cerebrospinal fluid. Their natural function as cell to cell communication vehicles makes them attractive for use as therapeutic shuttles to deliver biological molecules or drugs to target disease cells.

The XPack technology allows for cell-mediated generation of ready to use exosomes packed with any protein of choice. These exosomes can then be used to efficiently deliver proteins to target cells to alter or supplement biological pathways or be used to study exosome trafficking in vivo.

SBI has optimized a specific peptide sequence that targets a protein to the interior exosomal membrane, allowing the fusion protein to be packaged into exosomes for secretion. This “XPack” peptide sequence has been incorporated into the XPack cloning and expression lentivectors, allowing for reporter proteins or any protein of interest to be loaded into exosomes within a producer cell line. These engineered exosomes can then be isolated and delivered to target recipient cells. Additionally, stable XPack cell lines have been generated that serve as a constant source of exosomes loaded with reporter protein cargo, enabling tracking of exosome dynamics in both human cells and in vivo murine models. Purified, ready to use exosomes from these stable cell lines are also available as part of the comprehensive XPack system.


Technical References


Where can I find more information about SBI?

Visit the manufacturer page at www.stratech.co.uk/SBI, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


NEW Products: ProMag™ Particle Line Additions

ProMag™ from Bangs Laboratories are highly uniform polymer-based magnetic spheres available in diameters of 1µm and 3µm. A unique surface means low nonspecific binding in protein-based systems, and superior handling without the use of surfactant. These high-binding beads are suitable for use across a range of research and diagnostic applications, whether you’re working at laboratory scale or have the more stringent requirements of high throughput applications.


ProMag™ HC

NEW ProMag™ HC (High Capacity) 1μm is Bangs Laboratories latest advancement in magnetic particles. Intended for high capacity isolations, design parameters include high surface area and functionality.

Catalog Code Description Product Data Sheet Material Safety Data Sheet
PMC1HC ProMag™ HC 1 Series • COOH PDS 745 MSDS ProMag COOH-NH2
PMS1HC ProMag™ HC 1 Series • Streptavidin PDS 745 MSDS ProMag SA-GAM

ProMag™ HP

NEW ProMag™ HP (High Performance) have been meticulously engineered for use in assay development. The highly optimized composition ensures lowest autosignal, particularly with respect to chemiluminescence and exposed iron.

Catalog Code Description Product Data Sheet Material Safety Data Sheet
PMC3HP ProMag™ HP 3 Series • COOH PDS 743 MSDS ProMag COOH-NH2

Where can I find more information about Bangs Laboratories?

Visit the manufacturer page at www.stratech.co.uk/bangs_laboratories, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.


Product Focus: JC – 10 – Flow Cytometry Mitochondrial Membrane Potential Detection Kit

Key Benefits:

  • Superior solubility in aqueous buffers as compared to JC-1.
  • Applications – Cell permeable no wash Flow Cytometry assay.
  • Add reagent directly to live cells, incubate 30-60 minutes and measure.

Background

The mitochondrial permeability transition is an important step in the induction of cellular apoptosis. During this process, the electrochemical gradient (referred to as DY) across the mitochondrial membrane collapses. The collapse is thought to occur through the formation of pores in the mitochondria by dimerized Bax or activated Bid, Bak, or Bad proteins. Activation of these pro-apoptotic proteins is accompanied by the release of cytochrome c into the cytoplasm (1-4).

JC-1 has been used extensively to detect mitochondrial membrane potential, however due to its poor solubility in aqueous buffers it proves difficult to work with particularly when high concentrations are needed.

The JC-10 Assay Kit uses a unique cationic dye to signal the loss of the mitochondrial membrane potential. In healthy cells, the dye stains the mitochondria bright red. The negative charge established by the intact mitochondrial membrane potential allows the lipophilic dye, bearing a delocalized positive charge, to enter the mitochondrial matrix where it accumulates. When the critical concentration is exceeded, J-aggregates form which become fluorescent greenish orange where as it exists in the cytoplasm as monomers and fluoresces green. In apoptotic cells, the mitochondrial membrane potential collapses, and the JC-10 cannot accumulate within the mitochondria.

In these cells JC-10 remains in the cytoplasm in its monomeric green fluorescent form. Apoptotic cells, showing primarily green fluorescence, are easily differentiated from healthy cells which show greenish orange and green fluorescence. Both apoptotic and healthy cells can be visualized simultaneously by flow cytometry using 490 excitation and emission measured in FL1 (green) and FL2 (greenish orange). The JC-10 reagent is easy to use, simply dilute the reagent in the buffer provided or cell culture media and add it to the cells. After a 30-60- minute incubation analyze the samples via flow cytometry.

Fig A. Jurkat cells were cultured with DMSO for 2 hours. Cells were then stained with JC-10 Mitochondrial Membrane Potential Detection Kit for 15 minutes and analyzed by flow cytometry.

Fig B. Jurkat cells were cultured with staurosporine for 2 hours. Cell were then stained with JC-10 Mitochondrial Membrane Potential Detection Kit for 15 minutes and analyzed by flow cytometry.


Where can I find more information about Cell Technology?

Visit the manufacturer page at www.stratech.co.uk/cell_technology, email info@stratech.co.uk or call  +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.