Skip to main content

Proteome Profiler Human Pluripotent Stem Cell Array Kit

Contains 8 membranes - each spotted in duplicate with 15 different stem cell marker antibodies
siRNA-mediated Knockdown of PDGF R alpha, PDGF R beta, and cAbl Upregulates Expression of Pluripotent, Mesoderm, and Endoderm Markers.
(2)
Small Sample Lysate Volume Used to Detect Germ Layer Markers in Differentiated Human Pluripotent Stem Cells.
Catalog #
Availability
Size / Price
Qty
Loading...
ARY010
R&D Systems Guarantee

Product Performance Pledge

We offer a 100% guarantee on all our products.

Learn More »

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, the expression of stem cell markers is assessed using the following procedure:

  • Incubate cell lysates with the antibody-spotted array
  • Add biotinylated detection antibodies
  • Add streptavidin-HRP reagents
  • Detect signal by chemiluminescence
 

 

Reagents Provided

Reagents Supplied in the Proteome Profiler Human Pluripotent Stem Cell Antibody Array Kit (Catalog # ARY010)

  • 8-well Rectangular Multi-dish
  • Human Pluripotent Stem Cell Array - 8 nitrocellulose membranes each containing 15 different antibodies to stem cell markers printed in duplicate
  • Array Buffer 1
  • Array Buffer 2 Concentrate (5X)
  • Array Buffer 3
  • Chemi Reagent 1
  • Chemi Reagent 2
  • Detection Antibody Cocktail, Human Pluripotent Stem Cell Array
  • Lysis Buffer 16
  • Streptavidin-HRP
  • Transparency Overlay Template
  • Wash Buffer Concentrate (25X)
Other Supplies Required

Reagents

  • Aprotinin
  • Leupeptin (Catalog # 1167)
  • Pepstatin (Catalog # 1190)
  • Deionized or distilled water

Materials

  • Pipettes and pipette tips
  • Gloves
  • Plastic containers with the capacity to hold 50 mL (for washing the arrays)
  • Plastic transparent sheet protector (trimmed to 10 cm x 12 cm and open on three sides)
  • Plastic wrap
  • Paper towels
  • Absorbent lab wipes
  • Autoradiography cassette
  • X-ray film (Kodak® BioMax Light-1) or equivalent
  • Flat-tipped tweezers

Equipment

  • Rocking platform shaker
  • Microcentrifuge
  • Film developer
  • Flatbed scanner with transparency adapter capable of transmission mode

 

Procedure Overview

Add 1 mL of Array Buffer to each well of an 8-well multi-dish.

Add 1 mL of Array Buffer to each well of an 8-well multi-dish.

Place each membrane that will be used in one well of the 8-well multi-dish. The number on the membrane should be facing upward.

Incubate for one hour on a rocking platform shaker. Orient the tray so that each membrane rocks end to end in its well.

Place each membrane that will be used in one well of the 8-well multi-dish.

Replace the Array Buffer with prepared samples containing cell extract, Array Buffer 1, and Lysis Buffer for a final volume of 1 mL.

Incubate overnight at 2 °C to 8 °C on a rocking platform.

Wash each array 3 times with Wash Buffer in a separate container.

Wash each well of the 8-well multi-dish.

Replace the Array Buffer with prepared samples containing cell extract, Array Buffer 1, and Lysis Buffer for a final volume of 1 mL

Add 1 mL of diluted Biotinylated Detection Antibody Cocktail into each well.

Add the array to the well.

Incubate for 1 hour on a rocking platform.

Wash each array 3 times with Wash Buffer in a separate container.

Wash each well of the 8-well multi-dish.

Add 1 mL of diluted Biotinylated Detection Antibody Cocktail into each well

Add 1 mL of diluted Streptavidin-HRP to each well.

Add the array to the diluted Streptavidin-HRP.

Incubate for 30 minutes on a rocking platform.

Wash each membrane 3 times with Wash Buffer in a separate container.

Wash each well of the 8-well multi-dish.

Add 1 mL of diluted Streptavidin-HRP to each well

Place the array on a plastic sheet protector.

Add 0.5 mL of the prepared Chemi Reagent Mix evenly onto the array.

Cover the array with the top sheet of the plastic protector and incubate for 1 minute.

Blot off excess Chemi Reagent Mix.

Wrap the array and sheet protector in plastic wrap.

Place the wrapped array in an autoradiography film cassette and expose to film.

Place the array on a plastic sheet protector

Citations (19)

Showing  1 - 10 of 19 citations Showing All
Filter By:
Search Title:

Citations are publications that use Bio-Techne products. Selected citations for Proteome Profiler Human Pluripotent Stem Cell Array Kit include:

There are no citations that match your criteria.

Showing  1 - 10 of 19 citations Showing All

FAQs

  • Do I need to use the protease inhibitors (e.g. aprotinin, Ipegal) if I only plan to use cell supernatants?

    The proteinase inhibitors are recommended for cell and tissue lysates only.  You do not need them for cell culture supernates.

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Complete Your Research

Complete Your Experiment