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Proteome Profiler Human XL Oncology Array

Catalog # ARY026 | R&D Systems, Inc. a Bio-Techne Brand
Contains 4 membranes-each spotted in duplicate with 84 different cancer-related antibodies

Key Product Details

Proteome Profiler Human XL Oncology Array
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Setting the standard in quality research reagents

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, relative expression levels of human cancer-related proteins 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 Human XL Oncology Array nitrocellulose membranes spotted with 84 different antibodies to cancer-related proteins
  • Array Buffer 4
  • Array Buffer 6
  • Chemi Reagent 1
  • Chemi Reagent 2
  • Detection Antibody Cocktail, Human XL Oncology Array
  • Streptavidin-HRP
  • Transparency Overlay Template
  • Wash Buffer Concentrate (25X)

 

Other Supplies Required

Reagents

  • 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 17
  • Aprotinin
  • Leupeptin (Catalog # EI002)
  • Pepstatin (Catalog # EI003)

Other Supplies Required for Tissue Lysate Samples

  • Protease Inhibitor Cocktail
  • Igepal® CA-630
  • Sodium deoxycholate
  • Sodium dodecyl sulfate

 

Procedure Overview

R&D Systems Protocol for Multiple Analyte Detection Using the Proteome Profiler Human XL Oncology Array Kit, Panel A (Catalog # ARY026)

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

ARY026 Step 1

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

Incubate for one hour on a rocking platform shaker.

ARY026 Step 2

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.

ARY026 Step 3

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.

ARY026 Step 4

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

ARY026 Step 5

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

ARY026 Step 6

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.

ARY026 Step 7

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.

ARY026 Step 8

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.

ARY026 Step 9

Citations for Proteome Profiler Human XL Oncology Array (25)

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Customer Reviews for Proteome Profiler Human XL Oncology Array (7)

4.9 out of 5
7 customer ratings
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  • Proteome Profiler Human XL Oncology Array
    Name: Anonymous
    Verified Customer | Posted 12/06/2021
  • Proteome Profiler Human XL Oncology Array
    Name: Anonymous
    Verified Customer | Posted 03/25/2021
    Proteome Profiler Human XL Oncology Array ARY026
  • Proteome Profiler Human XL Oncology Array
    Name: Anonymous
    Verified Customer | Posted 01/29/2021
    Proteome Profiler Human XL Oncology Array ARY026
  • Proteome Profiler Human XL Oncology Array
    Name: Anonymous
    Verified Customer | Posted 04/20/2020
    This Oncology array kit is an excellent option do decide what proteins are regulated. Saves a lot of time and money. Extracellular vesicles derived from16HBE cells after LPS stimulation.
    Proteome Profiler Human XL Oncology Array ARY026
  • Proteome Profiler Human XL Oncology Array
    Name: Elad Prinz
    Verified Customer | Posted 03/05/2020
    Gives very useful information. used with MDA-MB-231 cell lysate.
    Proteome Profiler Human XL Oncology Array ARY026
  • Proteome Profiler Human XL Oncology Array
    Name: Anonymous
    Verified Customer | Posted 07/31/2018
    Proteome Profiler Human XL Oncology Array ARY026
  • Proteome Profiler Human XL Oncology Array
    Name: Anonymous
    Verified Customer | Posted 03/08/2017
    Proteome Profiler Human XL Oncology Array ARY026

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FAQs for Proteome Profiler Human XL Oncology Array

  • 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. 

  • Could you tell us whether this MMP-9 antibody can detect TIMP and MMP-9 complex?

    We haven't performed a large amount of specificity testing per each spot in the arrays because these are qualitative screening tools. However, we would recommend treating the MMP-9 spot as a "Total MMP-9" detecting both free MMP-9 and MMP-9 in complex. If the MMP-9 spot turns out to be of high interest for the customer's study, they will want to examine this target in more detail with quantitative tools such as ELISA or Luminex.

Product Documents for Proteome Profiler Human XL Oncology Array

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.