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Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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The STEMCCA Cre-Excisable Constitutive Polycistronic Lentivirus Kit contains high titer polycistronic lentivirus & Polybrene transfection reagent that have been validated for the generation of mouse & human induced pluripotent stem cells.
More>>The STEMCCA Cre-Excisable Constitutive Polycistronic Lentivirus Kit contains high titer polycistronic lentivirus & Polybrene transfection reagent that have been validated for the generation of mouse & human induced pluripotent stem cells. Less<<
SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
The STEMCCA Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Kit contains high titer polycistronic (OKSM) lentivirus and Polybrene transfection reagent that have been validated for the generation of mouse and human induced pluripotent stem cells from mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (HFFs), respectively. The Cre-excisable STEMCCA vector, comprised of the transcription factors Oct-4, Klf4, SOX-2, and c-Myc (OKSM) separated by the self-cleaving 2A peptide and IRES sequences, includes flanking LoxP sites that enable Cre-mediated excision of the vector following reprogramming (Sommer CA, 2009). The use of a single polycistronic lentiviral vector significantly improves reprogramming efficiencies and reduces the number of viral integrations. Furthermore, the ability to remove the reprogramming vector has been shown to improve the developmental potential of iPS cells and significantly increase their capacity to undergo directed differentiation in vitro (Sommer CA, 2010).
Important note: the STEMCCA lentivirus reprogramming kits are provided in two formats, dependent on whether you are working with a human or mouse system. Human reprogramming experiments require a higher multiplicity of infection (MOI) than mouse experiments. Therefore, the larger, three-vial kits are required for human reprogramming experiments. If you are reprogramming mouse embryo fibroblast cells, you have the option to use either the one- or three-vial kit, depending on how many wells you want to infect. Reprogramming other rodent somatic cells may require more than one vial of lentivirus to perform at optimal MOI.
References
Product Information
Components
1. EF1α-STEMCCA-LoxP (OKSM) Lentivirus: (Part number CS204439) One (1) vial containing 15 µL of high titer lentivirus.
2. Polybrene Transfection Reagent: (Part number TR-1003-50UL) One (1) vial containing 50 µL of 10 mg/mL stock.
The STEMCCA Cre-Excisable Constitutive Polycistronic Lentivirus Kit contains high titer polycistronic lentivirus & Polybrene transfection reagent that have been validated for the generation of mouse & human induced pluripotent stem cells.
Biological Information
Stem Cell Type
Mouse Embryonic Stem Cells
Induced Pluripotent Stem Cells
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Tested to confirm the generation of iPS cells from p3 mouse embryonic fibroblasts and p6 human foreskin fibroblasts. Cre-mediated excision of the reprogramming transgenes has only been validated in the mouse system. Other cell types have not been tested and thus similar results can not be guaranteed.
Usage Statement
This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges Merck to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Lentivirus is stable for at least 6 months when stored at -80°C. After first thaw, place immediately on ice and store in working aliquots to avoid further freeze thaws. Avoid freeze thaws as this will result in a decrease in the virus titer. Polybrene transfection reagent is stable for at least 1 year when stored at -20°C.
Excision of viral reprogramming cassettes by Cre protein transduction enables rapid, robust and efficient derivation of transgene-free human induced pluripotent stem cells. Kadari, Asifiqbal, et al. Stem Cell Res Ther, 5: 47 (2014)
2014
Integrating viruses represent robust tools for cellular reprogramming, however, the presence of viral transgenes in induced pluripotent stem cells (iPSCs) is deleterious as it holds the risk of insertional mutagenesis leading to malignant transformation. Here, we combine the robustness of lentiviral reprogramming with the efficacy of Cre recombinase protein transduction to derive iPSCs devoid of transgenes. By genome-wide analysis and targeted differentiation towards the cardiomyocyte lineage, we show that transgene-free iPSCs are superior to iPSCs before Cre transduction. Our study provides a simple, rapid and robust protocol for the generation of clinical-grade iPSCs suitable for disease modeling, tissue engineering and cell replacement therapies.
