NEW Supplier: SynVivo – Cell-Based Microchip Platform

Stratech Scientific now supply, SynVivo (CFD Research Corporation) products.

SynVivo products are a physiological, cell-based microchip platform that provides a morphologically and biologically realistic microenvironment allowing real-time study of cellular behavior, drug delivery and drug discovery.
SynVivo’s patented technology for recreating microvasculature scale, shape, fluidics, and cellular constructs provides the most realistic in vitro testing platform for drug discovery, delivery, efficacy, and toxicity assays.

Static well plate and linear flow channel assays cannot differentiate between shapes of drug delivery vehicles, whereas SynVivo can. As particle shape becomes a more relevant area of study for drug delivery, SynVivo can provide new insights that can help optimize drug delivery.

SynVivo can be used to capture real-time distribution of drugs in the microvasculature. Stagnant and recirculation zones that affect overall drug delivery and distribution can be readily identified.SynVivo can be used to quantify drug particle distribution patterns through analysis of shear rate and adhesion.

In addtion, SynVivo can support the growth of endothelial cells, cancer cells and many other cell types for mimicking healthy and disease conditions. In addition, targeted drug delivery can be quantified and optimized using the in vivo morphology and fluidic conditions in SynVivo.

For further information on available products visit our product listing.

Below is a video illustrating how the system works.


Basic Research

Cellular behavior and microcirculatory dysfunction:

SynVivo chips can be used to study real-time cell-cell interactions for:

  • Inflammation (leukocyte-endothelium),
  • Cancer (tumor-endothelium),
  • Thrombosis (platelet-endothelium), and
  • Infection (microbial-endothelium)

under physiological fluid flow conditions representing healthy and pathological conditions.

Leukocytes rolling and adhering on endothelial cells Infection of brain neuronal cells Infection on neuronal cells

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: EZ-TAL Effector Assembly Kit – Genome editing at your fingertips

Complete TALE assembly kit

  • Simple 2-day protocol
  • Assemble specific TALE-Nucleases
  • Build your own transcription activators

The easiest way to make your own custom TALE Nucleases and Transciption Activators
Transcription activator-like effectors (TALEs) are a class of naturally occurring transcription effectors that recognize specific DNA sequences to modulate gene expression. First identified in the plant pathogen Xanthomonas, the modularity of the TALE DNA binding domain enables sequence-specific perturbation of gene function and offers broad applications in genetic and epigenetic studies.

The TALE binding domain is defined by a simple, yet elegant code of tandem repeat sequences called repeat-variable di-residues (RVDs) that allow for the predictable generation of customized TALEs that bind to any DNA sequence being targeted with high levels of specificity. Compared to other gene targeting technologies such as zinc-finger nucleases (ZFNs) and meganucleases, TALEs offer the researcher more latitude in selecting potential DNA targets, ease in design, and provides levels of specificity on par or better than the aforementioned technologies.


EZ-TAL Kit Components and Assembly Workflow

Everything you need to make your own TALEs
Each EZ-TAL Kit contains the following components with enough material to perform a minimum of 6 and a maximum of 12 TALE assemblies, depending on the target sequence redundancy. The products are supplied in two boxes. Box 1 contains the entire TALE monomer library to built the multimers. Box 2 has all of the enzymes, buffers, substrates and colony PCR screening primers. Choose a genomic sequence target region and use the online software: TAL Effector Nucleotide Targeter website. Once your target sequences are selected, use the EZ-TAL ready-made monomer library to begin building your custom TAL Effector in just 2 days. The first step of the do-it-yourself protocol is to assembled monomers into multimers using a procedure based on restriction, ligation and amplification. Multimers 1 and 2 are designed to be hexamers, but the length of multimer 3 can vary to allow variations in the final length, such as 14-19 bp binding sequences. The correctly assembled circular multimers were subsequently amplified by PCR. On day 2, gel-purified multimers are assembled into a vector of choice, using a second restriction-ligation-based procedure, followed by bacterial transformation. Colony PCR is then performed for confirmation of insert size; typically, 40-90% of colonies displayed correct insert size. We recommend to use two colonies of correct insert size for sequence confirmation; typically 80-90% of sequences revealed correct assembly. The last step is to test your TAL Effectors for activity, the surveyor nuclease assay can be used to determine cutting efficiency at the desired genomic location. A schematic of the entire work flow is shown below.


Products

 

EZ-TAL Nuclease and Transcription factor Assembly Kits

EZ-TAL Assembly Kit-CMV-TALEN 1 Kit GE100A-1
EZ-TAL Assembly Kit-EF1a-TALEN 1 Kit GE120A-1
EZ-TAL Assembly Kit-MSCV-TALEN 1 Kit GE140A-1
EZ-TAL Assembly Kit-CMV-TALE-TF 1 Kit GE500A-1
EZ-TAL Assembly Kit-EF1a-TALE-TF 1 Kit GE520A-1
EZ-TAL Assembly Kit-MSCV-TALE-TF 1 Kit GE540A-1
EZ-TAL End-Monomer Kit (Contains end-monomers at positions 12, 13, 14, 15, 16, 17). Compatible with GE1xx-5xxA-1 (required) to generate TALEs with various lengths of 14 to 19-mers. 1 Kit provides a minimum of 6 TALEN assemblies. 1 Kit GE900A-1
EZ-TAL Multimer Amplification Primer Mix (10uM) 50 ul GE590PR-1
EZ-TAL Colony PCR Primer Mix (10uM) 50 ul GE595PR-1

 

For TALE-Nuclease assembly, SBI recommends Catalog # GE100A-1, as the CMV promoter is generally the strongest and most widely used option in the literature for TALEN applications.

 

TAL Effector Services

TALE Nuclease pair for sequence-specific??gene editing 10 ul plasmid DNA CS810TN-1
Artificial gene activator with a custom TALE DNA binding domain 10 ul plasmid DNA CS820TA-1
Artificial gene inhibitor with a custom TALE DNA binding domain 10 ul plasmid DNA CS820TR-1
A dual-reporter (GFP and luciferase) for monitoring TA/TR functionality 10 ul plasmid DNA CS830DR-1

Online Resources


Product Citations

  1. Uhde-Stone C, Sarkar N, Antes T, Otoc N, Kim Y, Jiang YJ, Lu B. A TALEN-based strategy for efficient bi-allelic miRNA ablation in human cells. RNA. 2014 Apr 9. [Epub ahead of print].
  2. Uhde-Stone C, Gor N, Chin T, Huang J, Lu B. A do-it-yourself protocol for simple transcription activator-like effector assembly. Biol Proced Online. 2013 Jan 14;15(1):3. doi: 10.1186/1480-9222-15-3.

Technical References

  1. Bedell, V. M., Y. Wang, J. M. Campbell, T. L. Poshusta, C. G. Starker, R. G. Krug Ii, W. Tan, S. G. Penheiter, A. C. Ma, A. Y. Leung, S. C. Fahrenkrug, D. F. Carlson, D. F. Voytas, K. J. Clark, J. J. Essner & S. C. Ekker (2012) In vivo genome editing using a high-efficiency TALEN system. Nature, doi: 10.1038/nature11537.
  2. Boch, J., H. Scholze, S. Schornack, A. Landgraf, S. Hahn, S. Kay, T. Lahaye, A. Nickstadt & U. Bonas (2009) Breaking the code of DNA binding specificity of TAL-type III effectors. Science, 326, 1509-12.
  3. Clark, K. J., D. F. Voytas & S. C. Ekker (2011) A TALE of two nucleases: gene targeting for the masses? Zebrafish, 8, 147-9. Doyle, E. L., N. J. Booher, D. S. Standage, D. F. Voytas, V. P. Brendel, J. K. Vandyk & A. J. Bogdanove (2012) TAL Effector-Nucleotide Targeter (TALE-NT) 2.0: tools for TAL effector design and target prediction. Nucleic Acids Res, 40, W117-22.
  4. Hockemeyer, D., H. Wang, S. Kiani, C. S. Lai, Q. Gao, J. P. Cassady, G. J. Cost, L. Zhang, Y. Santiago, J. C. Miller, B. Zeitler, J. M. Cherone, X. Meng, S. J. Hinkley, E. J. Rebar, P. D. Gregory, F. D. Urnov & R. Jaenisch (2011) Genetic engineering of human pluripotent cells using TALE nucleases. Nat Biotechnol, 29, 731-4.
  5. Huang, P., Z. Zhu, S. Lin & B. Zhang (2012) Reverse genetic approaches in zebrafish. J Genet Genomics, 39, 421-33. Lei, Y., X. Guo, Y. Liu, Y. Cao, Y. Deng, X. Chen, C. H. Cheng, I. B. Dawid, Y. Chen & H. Zhao (2012) Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs). Proc Natl Acad Sci U S A, PMID: 23045671.
  6. Li, L., M. J. Piatek, A. Atef, A. Piatek, A. Wibowo, X. Fang, J. S. Sabir, J. K. Zhu & M. M. Mahfouz (2012a) Rapid and highly efficient construction of TALE-based transcriptional regulators and nucleases for genome modification. Plant Mol Biol, 78, 407-16.
  7. Li, T., S. Huang, X. Zhao, D. A. Wright, S. Carpenter, M. H. Spalding, D. P. Weeks & B. Yang (2011) Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes. Nucleic Acids Res, 39, 6315-25. Li, T., B. Liu, M. H. Spalding, D. P. Weeks & B. Yang (2012b) High-efficiency TALEN-based gene editing produces disease-resistant rice. Nat Biotechnol, 30, 390-2.
  8. Liu, J., C. Li, Z. Yu, P. Huang, H. Wu, C. Wei, N. Zhu, Y. Shen, Y. Chen, B. Zhang, W. M. Deng & R. Jiao (2012) Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy. J Genet Genomics, 39, 209-15.
  9. Ma, S., S. Zhang, F. Wang, Y. Liu, Y. Liu, H. Xu, C. Liu, Y. Lin, P. Zhao & Q. Xia (2012) Highly Efficient and Specific Genome Editing in Silkworm Using Custom TALENs. PLoS One, 7, e45035.
  10. Mahfouz, M. M. & L. Li (2011) TALE nucleases and next generation GM crops. GM Crops, 2, 99-103.
  11. Mahfouz, M. M., L. Li, M. Piatek, X. Fang, H. Mansour, D. K. Bangarusamy & J. K. Zhu (2012) Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein. Plant Mol Biol, 78, 311-21.
  12. Moore, F. E., D. Reyon, J. D. Sander, S. A. Martinez, J. S. Blackburn, C. Khayter, C. L. Ramirez, J. K. Joung & D. M. Langenau (2012) Improved somatic mutagenesis in zebrafish using transcription activator-like effector nucleases (TALENs). PLoS One, 7, e37877.
  13. Mussolino, C. & T. Cathomen (2012) TALE nucleases: tailored genome engineering made easy. Curr Opin Biotechnol, 23, 644-50. Perez-Pinera, P., D. G. Ousterout & C. A. Gersbach (2012) Advances in targeted genome editing. Curr Opin Chem Biol, 16, 268-77.
  14. Reyon, D., S. Q. Tsai, C. Khayter, J. A. Foden, J. D. Sander & J. K. Joung (2012) FLASH assembly of TALENs for high-throughput genome editing. Nat Biotechnol. 30, 460-5.
  15. Uhde-Stone, C., J. Huang & B. Lu (2012) A robust dual reporter system to visualize and quantify gene expression mediated by transcription activator-like effectors. Biol Proced Online, 14, 8.

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.

 


Study Exosomes from Culture Media without FBS Contamination

Unless you are studying Cow exosomes, use Exo-FBS when isolating exosomes from cell culture media.

Exosomes isolated from cell culture media provide an exciting source for novel biomarker discovery and therapeutic potential. However, cells grown in FBS can have high levels of background from bovine serum, which typically contains an abundance of cow exosome vesicles. These cow exosomes can interfere with studies of exosomes secreted from cells of interest, for example, by contributing bovine microRNAs into a sample.

To improve research on tissue culture-derived exosomes, SBI has developed an exosome-depleted FBS growth supplement called Exo-FBS that has been stripped of bovine exosomes. Exo-FBS supports equivalent growth of many cell types in culture, is devoid of cow CD63 positive exosomes and does not have any measurable bovine microRNAs. Exo-FBS enables the study of cellular secreted exosomes in culture without the worry of contaminating cow exosomes. No ultracentrifugation is required. Exo-FBS is available as both a standard FBS supplement or heat-inactivated FBS media supplement (treated at 65°C for 15 minutes before bovine exosome removal).

HIGHLIGHTS

  • FBS with bovine exosome-sized vesicles removed
  • Stripped of CD63 positive cow exosomes
  • No detectable cow microRNAs
  • Same cellular growth rates supported as standard FBS
  • Use the same as standard FBS (add 10% in DMEM or RPMI)

Exo-FBS works with a broad range of cell types.


Sample Exo-FBS Product Citations:

Prostate Stromal Cells
Josson S, Gururajan M, Sung SY, Hu P, Shao C, Zhau HE, Liu C, Lichterman J, Duan P, Li Q, Rogatko A, Posadas EM, Haga CL, Chung LW. Stromal fibroblast-derived miR-409 promotes epithelial-to-mesenchymal transition and prostate tumorigenesis. Oncogene. 2014 Jul 28;0. doi: 10.1038/onc.2014.212.

Mesenchymal Stromal Cells
Xin H, Li Y, Liu Z, Wang X, Shang X, Cui Y, Gang Zhang Z, Chopp M. Mir-133b Promotes Neural Plasticity and Functional Recovery after Treatment of Stroke with Multipotent Mesenchymal Stromal Cells in Rats Via Transfer of Exosome-Enriched Extracellular Particles. Stem Cells. 2013 Apr 30. doi: 10.1002/stem.1409.

B-Cells
Momen-Heravi F, Bala S, Bukong T, Szabo G. Exosome-mediated delivery of functionally active miRNA-155 inhibitor to macrophages. Nanomedicine. 2014 Mar 29. pii: S1549-9634(14)00132-4.


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.


 

 

Product Focus: dNTPs & dNTP Pre-Mixes

Premium Quality dNTPs – at prices you can’t ignore…!

The synthesis of a complementary DNA/cDNA strand by enzymatic polymerization of deoxynucleotide triphosphates (dNTPs) is the basic principle of DNA amplification, sequencing and cDNA synthesis techniques such as PCR, real-time PCR and reverse transcription. For these, amplification sensitivity, product yield and reproducibility are strongly dependent on the quality of the reagents used.

Jena Bioscience’s enzymatic dNTP manufacturing process ensures superior quality for deoxynucleotide triphosphates: All their dNTPs are ultrapure (> 99 % by HPLC) and functionally tested by a set very sensitive enzymatic reactions (see below dNTP Guide and dNTP specifications).

Jena Bioscience dNTP Specifications


dNTP Mix / 10 mM

Premix of 10 mM dATP, dCTP, dGTP and dTTP

Product Cat. No. Amount
S pack NU-1006S-JEN 200 µl
L pack NU-1006L-JEN 1 ml

dNTP Mix / 25 mM

Premix of 25 mM dATP, dCTP, dGTP and dTTP

Product Cat. No. Amount
S pack NU-1023S-JEN 200 µl
L pack NU-1023L-JEN 1 ml

dNTP Mix incl. dUTP / 10 mM

Premix of 10 mM dATP, dCTP, dGTP and 20 mM dUTP

Product Cat. No. Amount
S pack NU-1020S-JEN 200 µl
L pack NU-1020L-JEN 1 ml

dNTP Mix GCamplifier / 10 mM

Modified dNTP Mix for amplification of GC-rich sequences

Product Cat. No. Amount
Kit PCR-257-JEN 100 µl

dNTP PCR Mix GCamplifier is specially designed for amplification of GC-rich DNA templates. The kit contains a modified dGTP analog that significantly reduces the stability of GC-rich sequences thereby enhancing PCR efficiency. The dNTP PCR Mix GCamplifier is used instead of a standard dNTP mix.


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.


 

Product Focus: Alexa Fluor® 680 & Alexa Fluor® 790 for Far-red and Infrared Detection on Western Blots

Alexa Fluor® 680 is a far-red-emitting dye with peak excitation at 684 nm and peak emission at 702 nm.  Alexa Fluor® 790 is an infrared-emitting dye with peak excitation at 783 nm and peak emission at 803 nm (Figure 1).  Antibodies conjugated with far-red- and Infrared-emitting dyes are more sensitive than those with dyes emitting visible light due to low fluorescence quenching of the conjugates, high extinction coefficients of the dyes, and low background autofluorescence.  The increased brightness allows for a wider range of immunofluorescence detection and imaging modalities.  Far-red and Infrared dye conjugates can be used for higher sensitivity Western blots, quantitative Western blots, in-gel Western blots, microWestern arrays, in-cell Western arrays, on-cell Western arrays, tissue section imaging, small animal whole body imaging, and other techniques that require the brightest dyes.

Figure 1. Excitation and emission spectra of Alexa Fluor® 680 (left) and Alexa Fluor® 790 (right) conjugated secondary antibodies.  All peaks were normalised.  Spectra were obtained with an M-Series spectrofluorometer from Photon Technology International, Inc. and an Ultraspec 1100 pro from Amersham Biosciences.

Jackson ImmunoResearch now offers the largest selection of Alexa Fluor® 680 and Alexa Fluor® 790 dyes conjugated with signal-enhancing primary antibodies, affinity-purified secondary antibodies, streptavidin, and purified IgG controls for single and double-labelling Western blots (Figure 2) and other tecniques requiring high sensitivity.  The secondary antibodies are adsorbed to eliminate cross-reactions with others species and with other immunoglobulin classes for double labelling.

 
Figure 2.  Double immunofluorescence staining on a Western blot using Alexa Fluor® 680 far-red dye and Alexa Fluor® 790 infrared dye.  Mouse IgG and goat IgG were reduced and denatured with ß-mercaptoethanol and SDS.  The heavy and light chains were separated by electrophoresis in SDS-PAGE, transferred to nitrocellulose, and double labelled with a 1:100,000 dilution of Alexa Fluor® 790-goat anti-mouse IgG, Fc? Subclass 1 specific (min X Hu, Bov, Rb Sr Prot, 115-655-205)(green) to detect heavy chains and a 1:100,000 dilution of Alex Fluor® 680-goat anti-mouse IgG, light chain specific (min X Bov, Gt, Hrs, Hu, Rb, Rat, Shp Ig, 115-625-174)(red) to detect light chains. Fluorescence was imaged in a LiCor Odyssey imager.  Goat IgG was used as a background control.  Note the faint bands of goat IgG heavy and light chains attesting to the extreme brightness of the dyes even at a dilution of 1:100000.

Both dyes can be used with LiCor Odyssey imaging systems and all the high-sensitivity techniques listed above.  We recommend that the antibodies be diluted at least 1:50,000 to 1:200,000 due to the high sensitivity of the conjugates.


Where can I find more information about Jackson ImmunoResearch?

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


Stratech Scientific’s Molecular Biology Product Range

Stratech Scientific supply Molecular Biology products from a range of different manufacturers, to give you all the reagents you need for your experiments in one place.  Additional information on this range can be found on the following pages:

Product Focus: Magic™ Mouse Adjuvant

KEY FEATURES

  • Highly effective — rapid production of high titers of antibodies. Only 2 immunizations are necessary.
  • Safe — non-toxic adjuvant with no adverse side effects to animals.
  • Immunogen and adjuvant-sparing — much less immunogen required than with conventional methods.
  • Time-saving — strong immune responses can be reached within 35 days after immunization.
  • Extremely valuable — in producing antibodies against conformational epitopes.

Magic™ Mouse Adjuvant is a novel immunization adjuvant specifically designed for rapid production of high titers of antibodies in mice. The adjuvant contains immune stimulatory CpG DNA — short oligonucleotides that contain unmethylated cytosine-guanine dinucleotides within a certain base context. The mammalian immune system has evolved to recognize these sequences, which are found naturally in bacterial DNA, as a sign of infection. Exposure to CpG DNA results in very rapid and strong immune activation, and, when applied with an antigen, CpG DNA produces high titers of antigen-specific antibodies. Different CpG DNA sequences activate the immune systems of different animal species. Magic™ Mouse Adjuvant contains a proprietary DNA sequence that is specifically designed for immunization of mice.

Among all research adjuvants and vaccine adjuvants available in the market, Magic™ Mouse Adjuvant by far is the most efficient mouse adjuvant for all types of immunogens tested, in terms of its rapidness in raising immune responses and the antibody titers it produces. Of note, this adjuvant does not yield comparable titers in other species including other types of rodents.

Importantly, by avoiding making water-in-oil emulsions, heating, sonification, lyophilization, or homogenization in immunogen preparation, which destroys the native conformation of the immunogen, this adjuvant allows production of antibodies against conformational epitopes on native antigens.


 KIT CONTENTS

CDN-A001-CRD 1.0 mL Magic™ Mouse Adjuvant, sufficient for immunization (priming and two boosts) of up to 10 mice.


Where can I find more information about Creative Diagnostics?

Visit the manufacturer page at www.stratech.co.uk/creative-diagnostic, 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.


 

STRATECH NEWS: Molecular Biology Special (Probe Issue 22)

Stratech now provide more than 800,000 products, and whilst our new website makes it easy to search for the products you know well, it can sometimes still be difficult to find related products in your area of
research.

Probe 22 is designed as a reference guide to help you find more of the Molecular Biology products that you are looking for, and maybe some that you didn’t know we offered.  It focuses on key products and groups of products, but also gives an overview of a greater range of products than previously.

More detailed information about individual products and techniques are available under the Areas section of the website under Molecular Biology and related sub areas:

The print version of Probe Issue 22 can be downloaded as a PDF.

To keep up to date with what is new and exciting at Stratech, visit our blog at http://blog.stratech.co.uk where you can search or use our predefined categories and tag clouds to browse related products.

Product Focus: Anti-mouse CD31 / DIA-310 Rat monoclonal anti-mouse endothelial cell marker CD31 (PECAM-1), Clone SZ31

Antibody clone SZ31 is the first antibody which reacts specifically with murine CD31 in formalin-fixed paraffin-embedded tissue sections.

CD31, also known as PECAM-1 (Platelet Endothelial Cell Adhesion Molecule-1) is expressed constitutively on the surface of embryonic and adult endothelial cells. It is also expressed on cell surfaces of monocytes, neutrophils, platelets and certain T-cell
subsets. It has been detected on bone marrow-derived hematopoetic stem cells and embryonic stem cells. CD31 is a 130kDa integral membrane glycoprotein and as a member of the immunoglobulin superfamily involved in the mediation of cell-to-cell adhesion.

CD31-mediated endothelial cell-cell interactions play a major role in angiogenesis. Studies have shown CD31 to be a superior marker in human angiogenesis, which reportedly predicts tumor recurrence. Pathophysiological studies of CD31 in murine model systems had limitations because standard formalin-fixed sections were excluded. The clone SZ31 eliminates these restrictions by allowing high quality immunohistochemical analysis of standard formalin-fixed paraffin sections in mice.


Product Information

MSDS        Datasheet

Catalogue numbers: DIA-310 (100μg)
DIA-310-M (20μg)
Specificity: CD31
Species Reactivity: Mouse
Host Species: Rat
Isotype: IgG2a
Clone: SZ31
Clonality (Mono-/Polyclonal): monoclonal
Application: Immunohistochemistry (IHC), Paraffin Sections (PS), Frozen Sections (GS), Western Blot
Conjugation: unconjugated
Format: lyophilisate, antibody purified (from culture supernatant)
Intended Use: for Research Use Only
Research Area: angiogenesis, CD markers, immunohistochemistry

Where can I find more information about Dianova?

Visit the manufacturer page at www.stratech.co.uk/dianova, 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: HR Targeting Vectors for CRISPR/Cas9 and TALE-based Editing

CRISPR/Cas9 and TALE-Nuclease technologies have provided researchers the ability to edit any genomic locus with site-specific DNA cutting. In order to fully leverage the power of these genome engineering nucleases, one can use homology-directed recombination for targeted insertions at DNA cleavage sites. For Knock-Out applications, using HR donor vectors enables positive selection of engineered cells, greatly reducing the amount of screening needed to isolate bi-allelic KO clones. For Gene Editing and Tagging applications, an HR template is required to introduce the desired nucleotide modifications or reporter tags. SBI has generated a suite of homologous recombination donor vectors to improve the efficiency of any genome engineering application.

Features include:

  • Simple HR arm additions flanking insulated expression cassettes
  • Built-in LoxP sites for downstream cassette removal
  • Detailed Application Notes providing step-by-step protocols
  • Live phone support from experienced genome engineering specialists
  • Custom HR Donor Design & Construction Services

How do I Knock-out and Knock-in genes using HR vectors?

For HR-based applications needing the disruption of a particular protein-coding or non-coding gene of interest, a gene knock-out targeting vector can be used. The HR process is initiated by site-specific DNA cleavage using either a CRISPR/Cas9 or TALE-Nuclease vector set within the region of interest. Next, simply clone 5′ and 3′ genomic sequences immediately flanking the targeted DNA sequence of interest to specifically direct the HR vector to insert an expression cassette into the targeted genomic location. This leads to the ‘knock-out’ of the sequence by HR cassette addition. The HR cassettes included in the PrecisionX vectors feature RFP, GFP and/or Puromycin/Hygromycin to facilitate the identification of cells that have undergone accurate genome engineering.

SBI Application Note for CRISPR-Cas9_Gene Knock-Out


How can I introduce single base changes into a target gene using HR vectors?

Learn how to introduce single base modifications into target genes for gene corrections and mutations. SBI provides HR vectors, CRISPR/Cas9 vectors and TALE-Nuclease systems to accomplish finely-targeted gene editing applications. Become a gene surgeon and create unique model cell lines with this powerful set of technologies.

SBI Application Note for CRISPR-Cas9 SBP Gene Editing


How can I tag endogenous genes for tracking using HR vectors?

Tag an endogenous sequence of interest with a marker to create a fusion or to co-express a marker tag with the endogenous sequence for various applications (e.g. tracking spatial dynamics of a protein, lineage tracking, etc.) with HR-tagging vectors. SBI provides a collection of four tagging and co-expression vectors for HR-based applications. Any type of DNA sequence, from protein-coding ORFs to non-coding sequences can be efficiently tagged or co-expressed with GFP and/or luciferase using these vectors. For protein-coding genes, the 3′ end (typically the last exon) can be targeted using these vectors for efficient generation of C-terminal fusion tagged proteins or co-expression of the gene and the marker tag via T2A or IRES elements.


Resources

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.