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Proteome Profiler Mouse XL Cytokine Array

Contains 4 membranes-each spotted in duplicate with 111 different cytokine antibodies
Detection of selected mouse cytokines in Balb/3T3 Cell Line Lysate
(3)
Detection of Selected Mouse Cytokines in CMT-93 Cell Line Lysate.
(3)
Detection of Selected Mouse Cytokines in Balb/3T3 Cell Line Supernate.
(3)
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Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, relative expression levels of mouse cytokines and chemokines in samples can be determined using the following procedure:

  • Prepare membrane and incubate with prepared sample
  • Incubate the membrane with Detection Antibody Cocktail
  • Incubate the membrane array with Streptavidin-HRP
  • Develop the membrane array with Chemi Reagents 1 and 2
  • Expose the membrane array to autoradiography film
 

 

Kit Contents
  • Rectangular 4-Well Multi-dish
  • 4 Mouse XL Cytokine Array nitrocellulose membranes spotted with 111 different antibodies to mouse cytokines
  • Array Buffer 4
  • Array Buffer 6
  • Chemi Reagent 1
  • Chemi Reagent 2
  • Detection Antibody Cocktail, Mouse XL Cytokine Array
  • Streptavidin-HRP
  • Transparency Overlay Template
  • Wash Buffer Concentrate (25X)

 

Other Supplies Required

Reagents

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

 

Procedure Overview

R&D Systems Protocol for Multiple Analyte Detection Using the Proteome Profiler Mouse XL Cytokine Array Kit, Panel A (Catalog # ARY028)

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

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.

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

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.

Add 0.5 mL Array Buffer 4 to each sample.

Replace the Array Buffer 6 in each well of the 4-Well Multi-dish with prepared samples.

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

Wash each array membrane 3 times with 1X Wash Buffer in a separate container.

Wash each well of the 4-Well Multi-dish with 1X Wash Buffer.

Add 0.5 mL Array Buffer 4 to each sample.
Add 30 µL of Detection Antibody Cocktail to 1.5 mL of Array Buffer 4/6 for each array.
Add 0.5 mL Array Buffer 4 to each sample.

Pipette 1.5 mL diluted Detection Antibody Cocktail into each well of the 4-Well Multi-dish.

Add 0.5 mL Array Buffer 4 to each sample.

Return each array membrane to the 4-Well Multi-dish containing diluted Detection Antibody Cocktail.

Incubate for one hour on a rocking platform shaker.

Wash each array membrane 3 times with 1X Wash Buffer in a separate container.

Wash each well of the 4-Well Multi-dish with 1X Wash Buffer.

Add 0.5 mL Array Buffer 4 to each sample.

Add 2 mL of diluted Streptavidin-HRP to each well of the 4-Well Multi-dish.

Place the array membrane in the diluted Streptavidin-HRP solution.

Incubate for 30 minutes on a rocking platform shaker.

Wash each array membrane 3 times with 1X Wash Buffer in a separate container.

Add 0.5 mL Array Buffer 4 to each sample.

Place the array membrane on a plastic sheet protector.

Pipette 1 mL of the prepared Chemi Reagent Mix evenly onto the membrane.

Cover the membrane with the top sheet of the plastic protector.

Incubate for 1 minute.

Blot off excess Chemi Reagent Mix.

Wrap the membrane and sheet protector in plastic wrap.

Place the wrapped array membrane in an autoradiography film cassette and expose to X-ray film.

Add 0.5 mL Array Buffer 4 to each sample.

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Customer Reviews (19)

4.8 out of 5
19 customer ratings
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  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 09/17/2020
    Excellent antibody array to screen a multitude of cytokines, chemokines, and growth factors in one small strip of membrane. Readout is not a fluorescence so you don't need a special fluorescence reader. If you are doing Western blotting, you can do this easily.
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 02/12/2020
    Mouse breast tumor tissues
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 01/25/2020
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 01/06/2020
    Excellent array!!
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 06/06/2019
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 03/13/2019
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 12/05/2018
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 11/11/2018
    Excellent specificity and detection for large number of cytokines. Works perfectly with little optimization.
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 11/05/2018
    Pancreas lysate. 750ug protein/ membrane
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 07/18/2018
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 04/01/2018
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 03/22/2018
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 12/07/2017
    200 uL pooled serum from wild-type C57Bl/6 mice.5 min exposure time
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 08/17/2017
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 07/17/2017
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 05/23/2017
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 11/28/2016
    150 uL of mouse serum
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 09/30/2016
    This is a great kit. It provides excellent reliability and a large number of cytokines, chemokines to screening in a single assay. I strongly recommend it.
  • Proteome Profiler Mouse XL Cytokine Array
    Name: Anonymous
    Verified Customer | Posted 08/26/2016
    The sample was cell culture supernatant collected from MEF.
    Proteome Profiler Mouse XL Cytokine Array Serum ARY028

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FAQs

  • Will the Array identification number stamped on the Array membrane interfere with detection if it is not cut-off before the membrane is blocked?

    The dye used for printing the Array identification number on the membranes will fluoresce and interfere with the LI-COR detection. It is critical that the number is cut off before beginning the experiment. 

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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