Proteome Profiler Pluripotent Stem Cell Array Kit, BioAssay™ (Human)

Analyzing the expression profiles of stem cell markers is helpful for understanding the differentiation mechanisms of human stem cells and developing disease treatments. The Human Proteome Profiler Pluripotent Stem Cell Array Kit, BioAssay™ is a rapid, sensitive, and economical tool to simultaneously detect the relative levels of expression of 15 stem cell markers without performing numerous individual immunoprecipitations and Western blots. Each antibody was carefully selected using cellular extracts prepared from cell lines known to express the target protein.

Key Benefits:
1. Verifies stem cell identity.
2. Detects 15 stem cell markers simultaneously.
3. Easier to perform than a Western blot.
4. Requires no specialized equipment.
5. The array protocol can now be performed with a smaller sample size. 6. Simultaneously detect pluripotency and germ layer markers using lysate from only 1 well of a 24-well plate.

Analytes Detected by this Kit:
Oct-3/4.
Nanog.
SOX2.
E-Cadherin.
Alpha-Fetoprotein (AFP).
GATA-4.
HNF-3 beta/FoxA2.
PDX-1/IPF1.
SOX17.
Otx2.
TP63/TP73L.
Goosecoid (Gsc).
Snail.
VEGF R2/KDR/Flk-1.
HCG.

Principle of the Assay:
Capture and control antibodies have been spotted in duplicate on nitrocellulose membranes. Cellular extracts are diluted and incubated overnight with the Human Pluripotent Stem Cell Array. The array is washed o remove unbound proteins, followed by incubation with a cocktail of biotinylated detection antibodies. Streptavidin-HRP and chemiluminescent detection reagents are applied, and a signal is produced at each capture spot corresponding to the amount of protein bound. Refer to the Appendix for a list and coordinates of analytes and controls.

Kit Components:
Human Pluripotent Stem Cell Array, 8 nitrocellulose membranes each containing 15 different anti-stem cell marker antibodies printed in duplicate.
Lysis Buffer 16: 21ml of a non-denaturing buffered solution.
Array Buffer 1: 21ml of a buffered protein base with preservatives.
Array Buffer 2 Concentrate (5X): 21ml of a concentrated buffered protein base with preservatives.
Array Buffer 3: 21ml of a buffered protein base with preservatives.
Wash Buffer Concentrate: 3 vials (21ml/vial) of a 25-fold concentrated solution of buffered surfactant with preservative. May turn yellow over time.
Detection Antibody Cocktail, Human Pluripotent Stem Cell Array: 1 vial of a biotinylated antibody cocktail; lyophilized
Streptavidin-HRP: 200ul
Chemi Reagent 1: 1 vial (2.5 mL)
Chemi Reagent 2: 1 vial (2.5 mL)
8-Well Rectangular Multi-dish: Clear 8-well rectangular multi-dish. Transparency Overlay Template: 1 transparency overlay template for coordinate reference.

Storage and Stability:
Unused kit is stable for 3 months at 4°C. Once opened may be stored for up to 3 months at 4°C; return unused membranes to the foil pouch containing the dessicant pack and reseal along entire edge of the zip seal.