3-Germ Layer Immunocytochemistry Kit

Catalog number: A25538

Molecular Probes™  Related applications: Cellular Imaging | Stem Cell Analysis

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Description

The 3-Germ Layer Immunocytochemistry Kit enables optimal image-based analysis of spontaneously differentiated embryoid bodies derived from human pluripotent stem cells. It detects widely accepted markers characteristic of the three embryonic germ layers: beta-III tubulin (TUJ1) for ectoderm, smooth muscle actin (SMA) for mesoderm, and alpha-fetoprotein (AFP) for endoderm. This high performance immunocytochemistry (ICC) kit includes a complete set of primary and secondary antibodies, a nuclear DNA stain, and premade buffers for an optimized staining experiment. To characterize undifferentiated pluripotent stem cells, we recommend use of the Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit.

With the 3-Germ Layer Immunocytochemistry Kit, you can:
• Confirm expression of common germ layer markers with highly specific antibodies
• Get more information per sample by measuring more than one marker at a time
• Generate beautiful multiplexed images with minimal effort
• Save time and avoid cell loss by minimizing unnecessary wash steps in your staining protocol
• Process samples with confidence using a complete and optimized set of ICC reagents

Confirm Expression of Common Germ Layer Markers
The marker antibodies included in the kit have been carefully selected and validated for high performance in the ICC analysis of the three germ lineages in randomly differentiated embryoid bodies. TUJ1, SMA and AFP are markers that are widely used for the detection of neurons (ectoderm), cardiomyocytes (mesoderm), and primitive hepatocytes (endoderm), respectively.

Get More Information Per Sample
To help save time and precious sample, the antibody combinations in the kit have been designed to allow for the specific and simultaneous assessment of two to three markers along with nuclear DNA staining, depending on the usage of the far red filter.

Generate Beautiful Multiplexed Images
Powered by the superior performance of bright, photostable Alexa Fluor® dyes, this kit enables the acquisition of beautiful multiplexed images using commonly available blue, green, orange/red, and far red filter sets.

Minimize Unnecessary Wash Steps
The streamlined method recommended for this kit eliminates unnecessary wash steps found in standard ICC protocols to help save time and decrease the likelihood of cell loss during the staining protocol, especially when using glass coverslips or chamber slides. The only washes required are after incubation with primary and secondary antibodies.

Process Samples with Confidence
An optimized set of premade fixation, permeabilization, blocking, and wash buffers allows you to focus on answering scientific questions rather than spending time preparing reagents and determining compatibility.

What You Get
Easily scaled to any culture plate, dish, or slide format, the 3-Germ Layer Immunocytochemistry Kit comes with reagents sufficient to stain 20 samples using a 200 µL staining volume. (See kit content details below.)
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Label or Dye: Alexa Fluor™ 647, 3 Color, Alexa Fluor™ 555, Alexa Fluor™ 594, Alexa Fluor™ 488
Product Size: 20 tests

Contents & storage

Store at -20°C to 4°C:
• 10 µL of Rabbit Anti-Human TUJ1 (500X)
• 40 µL of Mouse Anti-Human SMA IgG2a (100X)
• 10 µL of Mouse Anti-Human AFP IgG1 (500X)
• 20 µL of Alexa Fluor® 647 Donkey Anti-Rabbit (250X)
• 20 µL of Alexa Fluor® 488 Donkey Anti-Rabbit (250X)
• 20 µL of Alexa Fluor® 555 Goat Anti-Mouse IgG2a (250X)
• 20 µL of Alexa Fluor® 594 Goat Anti-Mouse IgG2a (250X)
• 20 µL of Alexa Fluor® 488 Goat Anti-Mouse IgG1 (250X)

Store at -20°C to RT:
• 1 vial of NucBlue® Fixed Cell ReadyProbes® Reagent (1X)
• 10 mL of Fixative Solution (1X)
• 10 mL of Permeabilization Solution S (1X)
• 20 mL of Blocking Solution (1X)
• 20 mL of Wash Buffer (10X)

Documents

Manuals & protocols

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