Proteome Profiler Human sReceptor Array, Hematopoietic

Refer to the product datasheet for complete product details.

View Graphic Procedure

• 2 Hematopoietic Array Membranes
• 2 Common Analyte Array Membranes
• Lysis Buffer 17
• Array Buffer 1
• Array Buffer 8
• Detection Antibody Cocktail H
• Detection Antibody Cocktail C
• Streptavidin-HRP
• Transparency Overlay Template
• Wash Buffer Concentrate (25X)

Reagents

• Aprotinin
• Leupeptin (Catalog # EI002)
• Pepstatin (Catalog # EI003)
• Igepal^® CA-630
• Deionized or distilled water

Materials

• Pipettes and pipette tips
• Gloves
• Plastic container 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
• Absorbent lab wipes (KimWipes^® or equivalent)
• Paper towels
• X-ray film (Kodak BioMax^™ Light-1) or equivalent
• Flat-tipped tweezers

Equipment

• Rocking platform shaker
• Microcentrifuge
• Autoradiography cassette
• Film developer
• Flatbed scanner with transparency adapter capable of transmission mode
• Computer capable of running image analysis software and Microsoft Excel

Other Supplies Required for Cell Lysate Samples

• Phosphate-Buffered Saline (PBS)
• Lysis buffer (1% Igepal CA-630, 20 mM Tris-HCl (pH 8.0), 137 mM NaCl, 10% glycerol, 2 mM EDTA, 10 µg/mL Aprotinin, 10 µg/mL Leupeptin, and 10 µg/mL Pepstatin)

Other Supplies Required for Tissue Lysate Samples

• PBS with protease inhibitors (10 µg/mL Aprotinin, 10 µg/mL Leupeptin, and 10 µg/mL Pepstatin)
• Triton^™ X-100

Add 2 mL of Array Buffer 6 to each well of the supplied 4-Well Multi-dish.

Place each array membrane in a separate well of the 4-Well Multi-dish.

Incubate for one hour on a rocking platform shaker.

Add 0.5 mL Array Buffer 4 to each sample.

Adjust volume of each sample to final volume of 1.5 mL with Array Buffer 6.