Human Three Germ Layer 3-Color Immunocytochemistry Kit

To analyze germ layer cells differentiated from human pluripotent stem cells.

Why Use Molecular Markers to Verify Human Stem Cell Pluripotency?

Research studies using embryonic stem (ES) or induced pluripotent stem (iPS) cells most often require weeks of cell culture before an experimental hypothesis can be tested.

Thus, the ability to verify pluripotency at an early stage in the experimental process is an important time saving step. Pluripotency status can be evaluated by the presence of stem cell markers and/or the ability of stem cells to differentiate into cells of the three germ layers. 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.

3-Color ICC using germ layer markers:

• Evaluates the pluripotency status of the starting stem cell population.
• Provides increased confidence in pluripotent status through the use of multiple markers for each germ layer.
• Ensures efficient use of time and reagents by utilizing single-step staining.
• Can be performed in randomly differentiated samples to simultaneously detect one marker from each germ layer.

This kit contains the following fluorochrome-conjugated antibodies to analyze human pluripotent stem cells differentiated into cells of each germ layer:

Ectoderm

• Anti-Human Otx2 NL557-Conjugated Goat IgG
• Anti-Human SOX1 NL493-Conjugated Goat IgG

Mesoderm

• Anti-Human Brachyury NL557-Conjugated Goat IgG
• Anti-Human HAND1 NL637-Conjugated Goat IgG

Endoderm

• Anti-Human GATA-4 NL493-Conjugated Goat IgG
• Anti-Human SOX17 NL637-Conjugated Goat IgG

Each antibody is supplied as a 10X solution; enough for 25 assays when used in a 50 µL staining volume per assay.

Stability and Storage

Reagents are stable for 12 months from date of receipt when stored in the dark at 2^° C to 8^°.

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 vitro and 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.