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Human/Mouse/Rat Neural Lineage Functional Identification Kit

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Verification of Neural Progenitor Cell Multipotency.
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SC028
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Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, NPC multipotency is verified using the following in vitro differentiation procedure:

  • Culture multipotent cells of interest
  • Induce astrocyte, neuron, and oligodendrocyte differentiation using a media supplement
  • Evaluate differentiation using mature phenotype marker antibodies and fluorescent ICC
 
 

Reagents Provided

Reagents supplied in the Human/Mouse/Rat Neural Progenitor Cell Functional Identification Kit (Catalog # SC028):

  • Neural Maintenance Supplement (500X)
  • Neural Differentiation Supplement (100X)
  • Bovine Fibronectin (100X)
  • NPC Marker: Goat Anti-Rat Nestin Antigen Affinity-purified Polyclonal Antibody
  • Astrocyte Marker: Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody
  • Neuron Marker: Mouse Anti-Neuron-specific beta-III Tubulin Monoclonal Antibody
  • Oligodendrocyte Marker: Mouse Anti-Oligodendrocyte Marker O4 Monoclonal Antibody

Note: The quantity of neural differentiation supplement in this kit is sufficient to make 50 mL of media for NPC expansion and 100 mL of medium for NPC differentiation. This is enough medium for the differentiation of two 24-well plates.

Other Supplies Required

Reagents

  • N-2 MAX Media Supplement (Catalog # AR009)
  • Poly-L-Ornithine
  • Phosphate buffered saline (PBS)
  • Penicillin-Streptomycin (100X)
  • Bovine serum albumin (BSA)
  • D-MEM/F-12 (1X)
  • Glucose
  • L-Glutamine
  • Sodium Bicarbonate (NaHCO3)
  • Trypan blue
  • 95% Ethanol
  • Deionized or distilled water
 

Materials

  • Human, mouse, or rat NPCs
  • 24-well culture plates
  • 12 mm coverslips
  • 15 mL centrifuge tubes
  • Pipettes and pipette tips
  • Serological pipettes
 

Equipment

  • 37 °C and 5% CO2 incubator
  • Centrifuge
  • Hemocytometer
  • Inverted microscope
  • 37 °C water bath
  • 0.2 µm filter unit, 250 mL
 

Procedure Overview

 
Prepare Poly-L-Ornithine- and Fibronectin-coated coverslips

Prepare Poly-L-Ornithine- and Fibronectin-coated coverslips.

 
 
Plate 0.5 -1.0 x 10<sup>6</sup> NPCs in media containing Neural Maintenance Supplement

Plate 0.5 -1.0 x 106 NPCs in media containing Neural Maintenance Supplement.

 

Culture cells to 50% confluency.

&nbsp;

 
Replace with fresh media after 24 hours

Replace with fresh media after 24 hours.

 

Culture cells for 48 hours after initial plating.

&nbsp;

 

ICC detection of Nestin to identify NPCs.

&nbsp;

 
Astrocyte Differentiation
Astrocyte
Differentiation
Neuron Differentiation
Neuron
Differentiation
Oligodendrocyte Differentiation
Oligodendrocyte
Differentiation
Day 1 Replace media with
Neural Differentiation Media.
Replace media with
Neural Differentiation Media.
Replace media with
Neural Differentiation Media.
Day 4 Repeat media change every 3 days. Repeat media change every 3 days. Repeat media change every 3 days.
Day 7-10 ICC detection of GFAP. ICC detection of beta-III Tubulin. ICC detection of Oligodendrocyte Marker O4.

 

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FAQs for Human/Mouse/Rat Neural Lineage Functional Identification Kit

  • Can Human/Mouse/Rat Neural Lineage Functional Identification Kit (Catalog # SC028) be used with both embryonic and adult neural progenitor cells?

    The Human/Mouse/Rat Neural Lineage Functional Identification Kit has been tested with embryonic neural progenitor cells. The kit should also work with adult neural progenitor cells, as adult cells have similar growth conditions.

Product Documents for Human/Mouse/Rat Neural Lineage Functional Identification Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.