Product Focus: Mycoplasma Detection Kit – QuickTest (10 min)

Stratech Scientific

All the steps can be done in the clean bench and the results will show up within 10 mins.

Should the cell culture system be contaminated by mycoplasma, typical metabolic enzymes from mycoplasma will degrade the culture medium components, with typical metabolite products secreted into the cell culture supernatant. The specificity of the metabolites produced by the mycoplasma are very high. Any other type of eukaryotic cells or bacteria will not produce the metabolites.
If the test sample contains the metabolites, the reaction system (including the test sample, the reaction sample well and the reaction buffer) will turn greenish-blue in color. The concentration of metabolites produced is proportional to how dark the color is, which directly indicates the amount of mycoplasma in each sample.

Biotool Mycoplasma Detection Kit – QuickTest

Advantages



Product Validation



Protocol



Where can I find more information about Biotool?

Visit the manufacturer page at www.stratech.co.uk/biotool, email

View original post 41 more words

SynVivo used to create the first neonatal blood-brain barrier model on a chip

Researchers at Temple University used the SynVivo® cell-based in vitro assay platform to model the attributes and functions of the neonatal stage blood-brain barrier (BBB) [1]. The SynVivo BBB model closely mimics the in vivo micro-environment including three-dimensional morphology, cellular interactions and flow characteristics on a microfluidic chip. This work marks the first dynamic in vitro neonatal BBB model that offers real time visualization and analysis and is suitable for studies of BBB function as well as screening of novel therapeutics.

“The work is important because studies of neonatal neuropathologies and development of appropriate therapeutics are hampered by a lack of relevant in vitro models of the neonatal blood-brain barrier,” said Dr. Sudhir Deosarkar, the lead author of this paper.

In the SynVivo-BBB model, which includes a tissue compartment and vascular channels placed side-by-side and separated by an engineered porous barrier, the researchers were able to co-culture neonatal rat brain endothelial cells and rat astrocytes under physiological conditions observed in vivo. The endothelial cells formed a full lumen and exhibited tight junction formation which increased under co-culture with astrocytes. The permeability of small molecules in the developed model was found to in excellent agreement with in vivo observations.

“The real-time visualization capabilities of the co-culture platform allowed, for the first time, visualization of astrocyte end-feet and endothelial cell interactions in an in vitro model,” said Prof. Mohammad Kiani who is the senior author of the paper. “This is a unique capability and will help us to understand and develop therapeutics for several developmental disorders and diseases of the brain.”


The PLOS ONE paper shows that in contrast to transwell models, the SynVivo based model exhibits significantly improved barrier characteristics similar to in vivo observations.


Reference

1A Novel Dynamic Neonatal Blood-Brain Barrier on a Chip. S. Deosarkar, B. Prabhakarpandian, B. Wang, J.B. Sheffield, B. Krynska, M. Kiani. PLOS ONE, 2015, DOI: 10.1371/journal.pone.0142725


Where can I find more information about SynVivo?

Visit the manufacturer page at www.stratech.co.uk/synvivo, 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: Annexin V-FITC Detection Kit

Annexin V-FITC kit provides a rapid and convenient assay for apoptosis.Annexin V-FITC kit detects the externalization of phosphatidylserine in apoptotic cells using recombinant annexin V conjugated to green-fluorescent fluorescein isothiocyante (FITC) dye and necrotic cells using propidium iodide (PI) by flow cytometry and fluorescence microscope. The AnnexinV-FITC kit uses annexin V conjugated with FITC to label phosphatidylserine sites on the membrane surface. The kit also includes propidium iodide (PI) stains necrotic cells with red fluorescence. After treatment with both probes, apoptotic cells show green fluorescence, dead cells show red and green fluorescence, and live cells show little or no fluorescence.

Biotool™ Annexin V-FITC Apoptosis Detection Kit Protocol


Where can I find more information about Biotool?

Visit the manufacturer page at www.stratech.co.uk/biotool, 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: Cell Meter™ Mitochondrial Hydroxyl Radical Detection Kit *Red Fluorescence*

The detection of intracellular hydroxyl radical is of central importance to understanding proper cellular redox regulation and the impact of its dysregulation on various pathologies. The hydroxyl radical (·OH) is one of the reactive oxygen species (ROS) highly reactive with other molecules to achieve stability. In general, hydroxyl radical is considered to be a harmful by-product of oxidative metabolism, which can cause molecular damage in living system. It shows an average lifetime of 10-9 nano seconds and can react with nearly every biomolecule such as nuclear DNA, mitochondrial DNA, proteins and membrane lipids.

AAT Bioquest’s Cell Meter™ Mitochondrial Hydroxyl Radical Detection Kit is optimized for detecting hydroxyl radical in mitochondria. MitoROS™ OH580 is live-cell permeant probe and can rapidly and selectively target hydroxyl radical in live cells. It generates red fluorescence when it reacts with ·OH, and can be easily read at Ex/Em= 540/590 nm. Cell Meter™ Mitochondrial Hydroxyl Radical Detection Kit provides a sensitive fluorimetric probe to detect OH· in live cells with one hour incubation. This kit can be used for fluorescence microplate readers and fluorescence microscopy applications.


Further Information

MiniCatalog: 1 2

Application Notes: 1 2


Where can I find more information about AAT Bioquest?

Visit the manufacturer page at www.stratech.co.uk/aatbioquest, 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: Ligands for P2 Receptors

Mono- and dinucleotide agonist and antagonist ligands for P2X and P2Y subtypes

Extracellular purine and pyrimidine nucleotides modulate the function of diverse mammalian cell types and tissues under both normal and pathophysiological conditions via corresponding purine and pyrimidine receptors such as the P2 receptor family.

Members of the P2 receptors can be divided into ligand-gated ion channels (P2X receptors) and G-protein-coupled receptors (GPCR, P2Y receptors), respectively (Figure 1).

Figure 1: Members of the P2 receptor superfamily.

Generally, P2-receptors have been linked to participation in several diseases such as cancer, cardiovascular, inflammatory and neuropathic diseases, including neuropathic pain. Thus, agonist and antagonist ligands for these receptors have a high potential for clinical applications.
They offer various mono- and dinucleotide agonist and antagonist ligands for functional studies of P2X and P2Y subtypes (see Table 1).
Table 1: Overview of P2Y and P2X receptor subtypes.

Ligands of P2Y Receptors Ligands of P2X Receptors
P2Y1 P2X1
P2Y2 P2X2
P2Y4 P2X3
P2Y6 P2X4
P2Y11 P2X5
P2Y12 P2X6
P2Y13 P2X7
P2Y14

Selected References

  1. Jacobson (2013) Structure based approaches to ligands for G-protein-coupled adenosine and P2Y receptors, from small molecules to nanoconjugates. J. Med. Chem. 56:3749.
  2. Volonte et al. (2009) Membrane components and purinergic signalling: the purinome, a complex interplay among ligands, degrading enzymes, receptors and transporters. FEBS J. 276:318.
  3. Jacobson et al. (2009) Development of selective agonists and antagonists of P2Y receptors. Purinergic Signalling 5:75.
  4. Jacobson et al. (2003) Engineering of A3 Adenosine and P2Y Nucleotide Receptors and their ligands. Drug Development Res. 58:330.
  5. Mueller (2002) P2-pyrimidinergic receptors and their ligands. Current Pharmaceutical Design 8:2353.
  6. Jacobson (2001) Probing adenosine and P2 receptors: design of novel purines and nonpurines as selective ligands. Drug Development Res. 52:178.

Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, 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: Viral & Bacterial Purified Proteins

Fitzgerald Industries have just launched a new line of viral and bacterial purified proteins. These proteins are produced in a native format and can be used for vaccination development and in a wide variety of serological based diagnostic assays.

Protein Type Size
Adenovirus type 40 protein Native 100ug/ 1mg
Adenovirus type 5 hexon protein Native 100ug/ 1mg
Astrovirus type 1 protein Native 200ul / 1ml
Bordetella pertussis FHA protein Native 100ug/ 1mg
Bordetella pertussis toxin protein Native 50ug / 100ug / 200ug / 1mg
Chlamydia trachomatis protein Native 1ml
Clostridium difficile Toxin A protein Native 2ug / 10ug / 50ug / 100ug / 500ug
Clostridium difficile Toxin B protein Native 2ug / 10ug / 50ug / 100ug
Clostridium difficile Toxoid A protein Native 50ug / 100ug / 500ug
Clostridium difficile Toxoid B protein Native 50ug / 100ug
CMV cell lysate protein Native 100ug/ 1mg
CMV protein Recombinant 100ug / 500ug / 1mg
CMV protein (High Purity) Native 100ug/ 1mg
Dengue NS1 protein (Serotype 1) Recombinant 100ug
Dengue NS1 protein (Serotype 2) Recombinant 100ug
Dengue NS1 protein (Serotype 3) Recombinant 100ug
Dengue NS1 protein (Serotype 4) Recombinant 100ug
Dengue VLP protein (Serotype 2) Recombinant 100ug
Dengue VLP protein (Serotype 3) Recombinant 100ug
Diphtheria Toxin protein Native 1mg
HSV1 lysate protein Native 1ml
HSV1 protein Native 200 ul / 1 ml
HSV2 lysate protein Native 1ml
HSV2 protein Native 200 ul / 1 ml
JEV NS1 protein Recombinant 100 ug / 500 ug
Mycoplasma pneumoniae protein Native 200 ug / 1 mg
Neisseria gonorrhoeae protein Native 1ml
TBE NS1 protein Recombinant 100 ug / 500 ug
Toxoplasma gondi IgG protein Native 200 ug / 1 mg
Toxoplasma gondi IgM protein Native 100 ug / 1 mg
Trichomonas vaginalis protein Native 1 ml
West Nile Virus NS1 protein Recombinant 100 ug
Yellow Fever Virus NS1 protein Recombinant 100 ug / 500 ug

Where can I find more information about Fitzgerald Industries International?

Visit the manufacturer page at www.stratech.co.uk/fitzgerald, 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.


 

Amplite™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Green Fluorescence

Hydrogen peroxide (H2O2) is a reactive oxygen metabolic by-product that serves as a key regulator for a number of oxidative stress-related states. It is involved in a number of biological events that have been linked to asthma, atherosclerosis, diabetic vasculopathy, osteoporosis, a number of neurodegenerative diseases and Down’s syndrome. Perhaps the most intriguing aspect of hydrogen peroxide biology is the recent report that antibodies have the capacity to convert molecular oxygen into hydrogen peroxide to contribute to the normal recognition and destruction processes of the immune system. Measurement of this reactive species will help to determine how oxidative stress modulates varied intracellular pathways. This Amplite™ Hydrogen Peroxide Assay Kit (11503-AAT) uses our unique Amplite™ Green hydrogen peroxide sensor to quantify hydrogen peroxide in live cells. Amplite™ Green is cell-permeable, and generates the green fluorescence when it reacts with hydrogen peroxide. The kit is an optimized ‘mix and read’ assay format that is compatible with HTS liquid handling instruments.



Further Information:   Protocol   References   MSDS   Spectrum  Catalogue


Where can I find more information about AAT Bioquest?

Visit the manufacturer page at www.stratech.co.uk/aatbioquest, 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: Dual CLICK-Labeling of Glycans

The dynamics of global glycan synthesis is an essential parameter to characterize the cellular response under various physiological and pathological conditions[1].

We now offer methylcyclopropene-functionalized derivatives of the natural monosaccharides N-Acetylmannosamine (Ac4ManCyoc) and N-Acetylglucosamine (Ac4GlcNCyoc) that are

  • incorporated instead of their natural counterpart thereby introducing a methylcyclopropene group into newly synthesized glycan structures of proteins[2-5].
  • ideally suited for dual labeling with Azide-functionalized monosaccharides[3,4] (Fig. 1) due to rapid visualization with Tetrazine-functionalized detection reagents via Cyclopropene-Tetrazine Ligation[5] which is compatible with Cu(I)-free strain-promoted Alkyne-Azide (SPAAC) labeling procedures.

Figure 1 – Azide- and Methylcyclopropene-functionalized monosaccharides facilitate dual glycan synthesis monitoring (modified according to [3]). Scale bar: 30 µM. HEK 293T cells were grown in the presence of 100 µM Ac4ManNCyoc & 50 µM Ac4GlcNAz (A), 100 µM Ac4ManNCyoc (B), 50 µM Ac4GlcNAz (C) and without a CLICKable monosaccharide (D). Nuclei were stained with Hoechst 33342 (blue). Azide-functionalized glycoproteins were visualized with 20 µM DIBO-488 via Cu(I)-free Strain-promoted Alkyne-Azide Click Chemistry (SPAAC). Methylcyclopropene-functionalized glycoproteins have been detected via Tetrazine-Cyclopropene ligation[5] with 25 µM Tetrazine-Cy3.

Selected References

  1. Laughlin et al. (2009) Imaging the glycome. Proc. Nat. Acad. Sci. 106 (1):12.
    Spaete et al. (2014) Expanding the scope of cyclopropene reporters for the detection of metabolically engineered glycoproteins by Diels-Alder reactions. Beilstein J. Org. Chem. 10:2235.
  2. Spaete et al. (2013) Rapid Labeling of Metabolically Engineered Cell-Surface Glycoconjugates with a Carbamate-Linked Cyclopropene Reporter. Bioconjugate Chem. 25:147.
  3. Patterson et al. (2014) Improved cyclopropene reporters for probing protein glycosylation. Mol. BioSyst. 10:1693.
  4. Patterson et al. (2012) Functionalized Cyclopropenes As Bioorthogonal Chemical Reporters. J. Am. Chem. Soc. 134:18638.

Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, 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.


 

Suppleir Focus: Avanti Polar Lipids NEW Products

Anandamide Phosphates

anandamide phosphate (AEA-P)

C20:4 anandamide phosphate (AEA-P) Avanti Number 870440

anandamide phosphate (OEA-P)

C18:1 anandamide phosphate (OEA-P) Avanti Number 870438

The phosphate ester of AEA, AEA-P, has been tested as a water soluble prodrug version of AEA in the treatment of C6 glioma cells in vivo. Here it acts with essentially the same potency as AEA.1 However, when tested for inhibition of AEA binding to isolated rat brain CB1 receptors, AEA-P is about 5-fold less potent as an agonist with a Ki of about 200nM.2 The phosphate esters of AEA and its analogs are also structural variants of LPA. However, the effects of AEA-P on the various LPA receptors have not been tested.

 

  1. Fowler, C.J., K.O. Jonsson, A. Andersson, J. Juntunen, T. Jarvinen, S. Vandevoorde, D.M. Lambert, J.C. Jerman, and D. Smart. (2003). Inhibition of C6 glioma cell proliferation by anandamide, 1-arachidonoylglycerol, and by a water soluble phosphate ester of anandamide: variability in response and involvement of arachidonic acid. Biochem Pharmacol 66:757-67.
  2. Sheskin, T., L. Hanus, J. Slager, Z. Vogel, and R. Mechoulam. (1997). Structural requirements for binding of anandamide-type compounds to the brain cannabinoid receptor. J Med Chem 40:659-67.

Interface inverse lipids with inorganic surfaces

(Δ9-Cis) PC (DOPC)

18:1 (Δ9-Cis) PC (DOPC) Avanti Number 850375

 

DOCP

DOCP Avanti Number 850311

DOCPe

DOCPe Avanti Number 850312

 

 

 

 

 

Zwitterionic phosphocholine (PC) lipids are highly biocompatible, representing a major component of the cell membrane. A simple mixing of PC liposomes and silica (SiO2) surface results in liposome fusion with the surface and formation of supported lipid bilayers. However, the stability of this bilayer is relatively low because adsorption is based mainly on weak van der Waals force. PC lipids strongly adsorb by TiO2 via chemical bonding with the lipid phosphate. The lack of fusion on TiO2 is attributable to the steric effect from the choline group in PC. In this study, inverse phosphocholine lipids (CP) are used, directly exposing the phosphate. Using a calcein leakage assay and cryo-TEM, fusion of CP liposome with TiO2 is demonstrated. The stability of this supported bilayer is significantly higher than that of the PC/SiO2 system, as indicated by washing the membrane under harsh conditions. Adsorption of CP liposomes by TiO2 is inhibited at high pH. Interestingly, the CP liposome cannot fuse with silica surface because of a strong charge repulsion. This study demonstrates an interesting interplay between a soft matter surface and metal oxides. By tuning the lipid structure, it is possible to rationally control the interaction force. This study provides an alternative system for forming stable supported bilayers on TiO2, and represents the first example of interfacing inverse lipids with inorganic surfaces.

  1. Wang, F. and J. Liu. (2015). A Stable Lipid/TiO Interface with Headgroup-Inversed
    Phosphocholine and a Comparison with SiO. J Am Chem Soc

Where can I find more information about Avanti Polar Lipids Inc.?

Visit the manufacturer page at www.stratech.co.uk/avanti-polar-lipids, 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: Enzyme-Assisted Terminal & Internal DNA CLICK Labelling

Reactive “CLICK”-groups such as azides, alkynes or DBCO are enzymatically introduced into DNA via modified nucleotides that are incorporated instead of their natural analogues. The resulting CLICK-functionalized DNA can subsequently be processed via CLICK Chemistry, offering the choice to introduce:

  • a biotin group for subsequent purification tasks (via Azides of Biotin, Alkynes of Biotin or DBCO-containing Biotinylation Reagents)
  • a fluorescent group for subsequent microscopic imaging (via Fluorescent Azides, Fluorescent Alkynes or DBCO-containing Fluorescent Dyes)
    or
  • to crosslink the DNA to complementary CLICK-functionalized biomolecules, such as proteins.

Winz et al.[1] recently reported a simple, modular and efficient strategy for enzymatic (terminal deoxynucleotidyl transferase – TdT) 3′-terminal labeling of DNA using nucleoside triphosphates analogs (NTPs) modified with “CLICKable” moieties. Further, they applied either ligation or primer extension to transform the 3′-terminal DNA-label into an internal one (Figure 1):

Figure 1 – Strategy for terminal or internal DNA labeling using terminal nucleotidyl transferase (TdT) and CLICK Chemistry according to Winz et al.[1]: (A) Incorporation of “CLICKable” nucleotides that contain Alkyne, Azide, DBCO or Norbornene moieties at the DNA 3′-end. (B) Transformation of 3′-labeled DNA into internally labeled DNA by ligation or primer extension. (C) “CLICK”-coupling of DNA to Biotin or Fluorophores.

Click Reagent Cat. No.
5-DBCO-dUTP CLK-T09
5-Ethynyl-dUTP (5-EdUTP) CLK-T07
5-Ethynyl-UTP (5-EUTP) CLK-T08
Azide-PEG4-aminoallyl-dUTP NU-1705
N6-(6-Azido)hexyl-3′-dATP NU-1707
N6-(6-Azido)hexyl-ATP CLK-NU-003
N6-(6-Azido)hexyl-dATP CLK-NU-002
N6-Propargyl-ATP (N6pATP) CLK-NU-001
Cy5-Azide CLK-CCA-9295

Reference:
[1] Winz et al. (2015) Nucleotidyl transferase assisted DNA labeling with different click chemistries. Nucleic Acids Research May 26. pii: gkv544.


Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, 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.

ki67 antibody