Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit

Catalog # Availability Size / Price Qty
SC024
Beta-III Tubulin, GFAP, and Oligodendrocyte Marker O4 in Differentiated Rat Cortical Stem Cells.
1 Image
Product Details
Procedure
FAQs
Supplemental Products
Reviews

Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit Summary

Key Benefits

  • Verifies multipotency to reduce experimental variation
  • Labels neurons, astrocytes, and oligodendrocytes
  • Can be used for simultaneous 3-color staining
 

 

Why Analyze Neural Progenitor Cell Proliferation and Differentiation In Vitro?

Research studies using neural progenitor cells (NPCs) most often require weeks of cell culture before an experimental hypothesis can be tested.

Thus, the ability to verify multipotency at an early stage in the experimental process is an important time saving step. Multipotency status can be evaluated by the presence of cell markers and/or the ability of progenitor cells to differentiate into multiple neural lineages. To determine if a cell population truly consists of multipotent NPCs, it is important to verify their ability to differentiate into neurons, astrocytes, and oligodendrocytes.

3-color ICC using neural cell markers:

  • Evaluates the multipotency status of the starting NPC population.
  • Increases confidence in multipotency status by combining 3 different fluorochrome-conjugated antibodies for simultaneous staining.
  • Reduces variability within an experiment and between studies.
  • Ensures efficient use of time and reagents by utilizing single-step staining.
  • Can be performed using randomly differentiated samples.
 

 

Kit Components

This kit contains the following fluorochrome-conjugated antibodies to verify neural stem cell multipotency:

  • NL557-conjugated Mouse anti-Human Oligodendrocyte Marker O4
  • NL637-conjugated Mouse anti-beta III Tubulin
  • NL493-conjugated Sheep anti-Human GFAP

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

 

 

Specifications

Source
N/A
Shipping Conditions
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Species
Human

Product Datasheets

You must select a language.

x

Scientific Data

Immunocytochemistry Beta-III Tubulin, GFAP, and Oligodendrocyte Marker O4 in Differentiated Rat Cortical Stem Cells. View Larger

Beta-III Tubulin, GFAP, and Oligodendrocyte Marker O4 in Differentiated Rat Cortical Stem Cells. Beta-III Tubulin, GFAP, and Oligodendrocyte Marker O4 were simultaneously detected in immersion-fixed 7 day differentiated Rat Cortical Stem Cells (Catalog # NSC001) using the Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit (Catalog # SC024). Proteins were detected using antibodies included in the kit: neurons were stained with a NorthernLights(NL)637-conjugated Mouse Anti-neuron Specific beta-III Tubulin Monoclonal Antibody (pseudocolored gray), astrocytes were stained with a NL493-conjugated Sheep Anti-GFAP Antigen Affinity-purified Polyclonal Antibody (green), and oligodendrocytes were stained with a NL557-conjugated Mouse Anti-Oligodendrocyte Marker O4 Monoclonal Antibody (red). The nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, neural stem cell (NSC) multipotency can be verified via 3-Color ICC using the following protocol:

  • NSCs are cultured on coated coverslips
  • Fluorochrome-conjugated primary antibodies are added to fixed cells
  • Neural cell markers are visualized using a fluorescence microscope
 

 

Reagents Provided

Reagents supplied in the Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit (Catalog # SC024):

  • NL557-conjugated Mouse anti-Human Oligodendrocyte Marker O4
  • NL637-conjugated Mouse anti-beta III Tubulin
  • NL493-conjugated Sheep anti-Human GFAP

 

Other Supplies Required

Reagents

  • Appropriate NSC culture substrate (e.g., Human Fibronectin (Catalog # 1918-FN-02M), Bovine Fibronectin (Catalog # 1030-FN), Poly-L-ornithine, etc.)
  • Cell culture medium (e.g., DMEM and N-2 Plus Media Supplement (Catalog # AR003) or equivalent)
  • 4% paraformaldehyde in PBS
  • 1% BSA in PBS
  • 0.3% Triton X-100, 1% BSA, 10% normal donkey serum in PBS
  • Mounting medium (Catalog # CTS011 or equivalent)

Materials

  • Differentiated NPCs
  • Coverslips (sterilized)
  • Cell culture plate (24-well)

Equipment

  • 37 °C and 5% CO2 incubator
  • Centrifuge
  • Hemocytometer
  • Inverted microscope
  • Fluorescence microplate

 

Procedure Overview

These antibodies have been tested for immunocytochemistry staining using 7 day differentiated rat cortical stem cells.

Coat coverslips with a NSC culture substrate.

Coat coverslips with a NSC culture substrate

Plate NSCs

Culture to desired confluency/age

Plate NSCs

Fix differentiated cells with 4% paraformaldehyde.

Fix differentiated cells with 4% paraformaldehyde

Block in blocking solution.

Block in blocking solution

Incubate with fluorochrome-conjugated primary antibodies.

Wash with wash buffer.

Incubate with fluorochrome-conjugated primary antibodies

Incubate with nuclear counterstain.

Incubate with nuclear counterstain

48 hours Mount the coverslip.

Incubate Visualize using a fluorescence microscope and appropriate filter sets.

48 hours Mount the coverslip

FAQs

No product specific FAQs exist for this product, however you may

View all Stem Cell Product FAQs
Loading...

Reviews for Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit

There are currently no reviews for this product. Be the first to review Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit and earn rewards!

Have you used Human/Mouse/Rat Neural 3-Color Immunocytochemistry Kit?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review