Description
General description
Mouse embryonic stem (mES) cells are pluripotent cells derived from the inner cell mass of pre-implantation blastocysts. Mouse induced pluripotent (miPS) cells are pluripotent cells generated by reprogramming mouse somatic cells using four transcription factors, Oct-4, Klf-4, Sox-2, and c-Myc, or their variants. Both mESC and miPSC can self-renew and have the ability to generate all three germ layers: ectoderm, mesoderm, and endoderm. In vitro, mESC/iPSC are normally maintained and propagated on mouse fibroblast feeders or in feeder-free conditions in media containing LIF/ESGRO. However, spontaneous differentiation may occur in subpopulations of cells. Several pluripotent markers are commonly used to distinguish pluripotent mESC/iPSC from differentiated cells.
EMD Millipore’s Fluorescent Mouse ES/iPS Cell Characterization Kit contains a range of sensitive tools for the phenotypic assessment of the pluripotent status of mouse ES/iPS cells. Included in the kit is an enzymatic assay to measure alkaline phosphatase activity in the cells along with validated directly conjugated antibodies to pluripotent transcription factors, Oct-4, Sox-2 and DPPA-2 and cell surface epitope SSEA-1 to enable rapid immunocytochemical marker analysis. The Dapi nuclear dye is conveniently included to aid in cell quantification. While the expression levels of pluripotent markers are expected to be diminished upon differentiation, each possess specific expression kinetics.
Application
Research Category
Stem Cell Research
Research Sub Category
Pluripotent & Early Differentiation
This Fluorescent Mouse ES/iPS Cell Characterization Kit contains a range of sensitive tools for the phenotypic assessment of the pluripotent status of mouse Embryonic stem & induced pluripotent Stem cells.
Biochem/physiol Actions
Cross Reactivty
None
Physical form
Purified conjugated mouse monoclonal antibodies are stored in PBS with 0.1% sodium azide and 15mg/ml BSA.
Preparation Note
Store at 2-8°C until expiration date of kit. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the expiration date.
Analysis Note
Conjugated antibodies and alkaline phosphatase reagents have been validated on mouse ES and STEMCCA derived mouse ips cells.
Other Notes
1. Fast Red Violet solution (Part No. 90239): One 15 mL bottle.
2. Napthol AS-BI phosphate solution (2mg/mL) in a buffer, pH 8.5 (Part No. CS235583). One 15 mL bottle.
3. Mouse anti-Oct-4 (POU5f1), clone 7F9.2, Alexa Fluor™ 488 conjugate (Part No. MAB4419A4-50UL). One vial containing 50 µL of 0.5 mg/mL conjugated monoclonal antibody.
4. Mouse anti-Sox-2, clone 10H9.1, Cy3 conjugate (Part No. MAB4423C3-50UL). One vial containing 50 µL of 0.5 mg/mL conjugated monoclonal antibody.
5. Mouse anti-DPPA-2, clone 6C1.2, Alexa Fluor 488 conjugate (Part No. MAB4356A4-50UL). One vial containing 50 µL of 0.5 mg/mL conjugated monoclonal antibody.
5. Mouse anti-SSEA-1, clone MC-480, Cy3 conjugate (Part No. MAB4301C3-50UL). One vial containing 50 µL of 0.5 mg/mL conjugated monoclonal antibody.
8. DAPI, 100 µL (Part No. 90229). One vial containing 100 µL volume.
Legal Information
ALEXA FLUOR is a trademark of Life Technologies
