产品介绍
Kit Summary
Verifies the pluripotency of your starting population
Only takes 5 days
Reduces experimental variation
To determine if a cell is truly a pluripotent stem cell, it is important to verify its ability to differentiate into each of the three germ layers.
The teratoma assay is a standard way to assess pluripotency, but this method requires expensive animal models and is time-consuming, often requiring 6 to 10 weeks before pluripotency status can be determined. Similarly, embryoid body assays are time-consuming and rely on random differentiation.
Uses fully defined supplements to drive reproducible differentiation.
Provides results in 5 days to save time and reagents.
Avoids tissue embedding and sectioning procedures.
Can verify the ability to differentiate into 3 germ layers without the use of an animal model.
Verifies a healthy, pluripotent starting stem cell population to increase consistency between studies and reduce unwanted experimental variability.
Kit Contents
This kit contains the following specially formulated media supplements and growth factors to drive pluripotent stem cell differentiation and a marker to characterize each of the three germ layer cell types.
Differentiation Base Media Supplement (50X)
Ectoderm, Mesoderm, and Endoderm Differentiation Supplements
Ectoderm Marker: Goat Anti-Human Otx2 Antigen Affinity-purified Polyclonal Antibody
Mesoderm Marker: Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal AntibodyV
Endoderm Marker: Goat Anti-Human SOX17 Antigen Affinity-purified Polyclonal Antibody
The quantity of each component in this kit is sufficient to make 200 mL of media for differentiation. This is enough media for the differentiation of one 24-well plate of each cell type.
The acute and chronic effects of over-exposure to the reagents in this kit are unknown. Safe laboratory handling procedures should be followed and protective clothing should be worn when handling kit reagents.
Data Examples
Verification of Induced Pluripotent Stem Cell Pluripotency.iPS2 human induced pluripotent stem cells were differentiated to ectoderm, mesoderm, and endoderm using the media supplements included in the Human Pluripotent Stem Cell Functional Identification Kit (Catalog # SC027). The kit also contains Goat Anti-Human Otx2 (ectoderm), Goat Anti-Human Brachyury (mesoderm), and Goat Anti-Human SOX17 (endoderm) Antigen Affinity-purified Polyclonal Antibodies for the confirmation of differentiation status. To further evaluate lineage commitment, cells were stained with a Goat Anti-Human SOX1 Antigen Affinity-purified Antibody (Catalog # AF3369), a Goat Anti-Human HAND1 Antigen Affinity-purified Antibody (Catalog # AF3168), and a Goat Anti-Human HNF-3 beta/FoxA2 Antigen Affinity-purified Antibody (Catalog # AF2400). The cells were stained using the NorthernLights™ 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001; red) and the nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Verification of Human Induced Pluripotent Stem Cell Pluripotency.Two human induced pluripotent stem cells lines (ADLF1 and FAB2) were differentiated to ectoderm, mesoderm, and endoderm using the media supplements included in the Human Pluripotent Stem Cell Functional Identification Kit (Catalog # SC027B). The kit also contains Goat Anti-Human Otx2 (ectoderm), Goat Anti-Human Brachyury (mesoderm), and Goat Anti-Human SOX17 (endoderm) Antigen Affinity-purified Polyclonal Antibodies for the confirmation of differentiation status. The cells were stained using the NorthernLights™ 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001; red) and the nuclei were counterstained with DAPI (blue). |
Preparation and Storage
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Background: Pluripotent Stem Cells
Embryonic stem (ES) cells are pluripotent stem cells derived from the inner cell mass of pre-implantation embryos. Induced pluripotent stem (iPS) cells can be generated by somatic cell reprogramming following the exogenous expression of specific transcription factors (Oct-3/4, KLF4, SOX2, and c-Myc). These cell types are capable of unlimited, undifferentiated proliferation in vitroand still maintain the capacity to differentiate into a wide variety of somatic cells. In this capacity, pluripotent stem cells have widespread clinical potential for the treatments of heart disease, diabetes, spinal cord injury, and a variety of neurodegenerative disorders.
R&D Systems offers a wide range of products to support pluripotent stem cell culture and differentiation. Mouse embryonic fibroblasts may be used to maintain and expand pluripotent stem cells in an undifferentiated state. We also offer defined culture media, which are specifically optimized for use with human or rodent pluripotent stem cells. In addition, R&D Systems offers a variety of products to assess differentiation status and identify specific stem cell types of interest, including panels of marker antibodies, primer pairs, multi-color flow cytometry kits, and specialized verification kits.
公司简介
R&D Systems作为全球免疫学和细胞生物学产品的领跑者,1976年创立于美国明尼苏达州。其母公司Bio-Techne(原Techne公司)于1983年在美国NASDAQ上市。
目前,R&D Systems拥有近30,000种产品,95%以上由自己研发生产。其产品包括细胞因子、ELISA试剂盒、生长因子、趋化因子、抗体、Animal-Free™蛋白、Luminex 液相芯片、多因子检测固相芯片、流式细胞分析与细胞筛选、干细胞及细胞培养等;覆盖肿瘤、发育、糖生物、内分泌、免疫、神经学、蛋白酶、信号传导等学科研究领域;且每年有近2,000种新产品不断问世,以满足不同科研工作者的研究需要。
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