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Proteome Profiler Human XL Oncology Array
R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
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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.
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.
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 30 µL of Detection Antibody Cocktail to 1.5 mL of Array Buffer 4/6 for each array.
Pipette 1.5 mL diluted Detection Antibody Cocktail into each well of the 4-Well Multi-dish.
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 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.
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.
Citations for Proteome Profiler Human XL Oncology Array (20)
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Citations are publications that use Bio-Techne products. Selected citations for Proteome Profiler Human XL Oncology Array include:
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Species: Human
Sample Types: Cell Lysates
M Zerfaoui et al. (2022), Nuclear Localization of BRAFV600E Is Associated with HMOX-1 Upregulation and Aggressive Behavior of Melanoma Cells Cancers, 14(2).
PMID: 35053476 -
Species: Human
Sample Types: Tissue Homogenates
CS Pulford et al. (2022), A Hybrid Epithelial to Mesenchymal Transition in Ex Vivo Cutaneous Squamous Cell Carcinoma Tissues International Journal of Molecular Sciences, 23(16).
PMID: 36012449 -
Species: Human
Sample Types: Cell Lysates
AM Alnuqaydan et al. (2022), Evaluation of the Cytotoxic Activity and Anti-Migratory Effect of Berberine-Phytantriol Liquid Crystalline Nanoparticle Formulation on Non-Small-Cell Lung Cancer In Vitro Pharmaceutics, 14(6).
PMID: 35745691 -
Species: Human
Sample Types: Cell Culture Supernates
A Anchan et al. (2022), Melanoma Mediated Disruption of Brain Endothelial Barrier Integrity Is Not Prevented by the Inhibition of Matrix Metalloproteinases and Proteases Biosensors, 12(8).
PMID: 36005056 -
Species: Human
Sample Types: Cell Lysates
Y Katsuda et al. (2022), Small molecule-based detection of non-canonical RNA G-quadruplex structures that modulate protein translation Nucleic Acids Research, 0(0).
PMID: 35801908 -
Species: Human
Sample Types: Cell Culture Supernates
I Fatima et al. (2021), MASTL regulates EGFR signaling to impact pancreatic cancer progression Oncogene, 0(0).
PMID: 34331012 -
Species: Human
Sample Types: Cell Culture Supernates
N Stoup et al. (2021), The EGF Domains of MUC4 Oncomucin Mediate HER2 Binding Affinity and Promote Pancreatic Cancer Cell Tumorigenesis Cancers, 13(22).
PMID: 34830899 -
Species: Human
Sample Types: Tissue Homogenates
Z Ni et al. (2021), Transcriptomic identification of HBx-associated hub genes in hepatocellular carcinoma PeerJ, 9(0):e12697.
PMID: 35036167 -
Species: Human
Sample Types: Protein
G Smagurausk et al. (2020), New Paradigms to Assess Consequences of Long-Term, Low-Dose Curcumin Exposure in Lung Cancer Cells Molecules, 25(2).
PMID: 31963196 -
Species: Human
Sample Types: Cell Culture Lysates
M Mosley et al. (2020), Immuno-imaging of ICAM-1 in tumours by SPECT Nucl. Med. Biol., 84(0):73-79.
PMID: 32135474 -
Species: Human
Sample Types: Cell Lysates
ES Grassi et al. (2020), Hypoxia-induced release, nuclear translocation, and signaling activity of a DLK1 intracellular fragment in glioma Oncogene, 0(0).
PMID: 32205867 -
Species: Human
Sample Types: Cell Lysates
A Argentiero et al. (2019), Gene Expression Comparison between the Lymph Node-Positive and -Negative Reveals a Peculiar Immune Microenvironment Signature and a Theranostic Role for WNT Targeting in Pancreatic Ductal Adenocarcinoma: A Pilot Study Cancers (Basel), 11(7).
PMID: 31277479 -
Species: Human
Sample Types: Cell Lysates
A Argentiero et al. (2019), Gene Expression Comparison between the Lymph Node-Positive and -Negative Reveals a Peculiar Immune Microenvironment Signature and a Theranostic Role for WNT Targeting in Pancreatic Ductal Adenocarcinoma: A Pilot Study Cancers (Basel), 11(7).
PMID: 31277479 -
Species: Mouse
Sample Types: Tissue Lysates
BT Elie et al. (2019), Preclinical evaluation of an unconventional ruthenium-gold-based chemotherapeutic: RANCE-1, in clear cell renal cell carcinoma Cancer Med, 0(0).
PMID: 31192543 -
Species: Human
Sample Types: Cell Lysates
SB Uppada et al. (2018), MASTL induces Colon Cancer progression and Chemoresistance by promoting Wnt/?-catenin signaling Mol. Cancer, 17(1):111.
PMID: 30068336 -
Species: Human
Sample Types: Cell Lysates
BM Ku et al. (2018), Acquired resistance to AZD9291 as an upfront treatment is dependent on ERK signaling in a preclinical model PLoS ONE, 13(4):e0194730.
PMID: 29641535 -
Species: Human
Sample Types: Cell Lysates
NM Scheers et al. (2018), Ferric citrate and ferric EDTA but not ferrous sulfate drive amphiregulin-mediated activation of the MAP kinase ERK in gut epithelial cancer cells Oncotarget, 9(24):17066-17077.
PMID: 29682205 -
Species: Human
Sample Types: Cell Lysates
G Hamilton et al. (2016), Small cell lung cancer: Circulating tumor cells of extended stage patients express a mesenchymal-epithelial transition phenotype Cell Adh Migr, 10(4):360-7.
PMID: 26919626 -
Species: Human
Sample Types: Cell Lysates
(2016), CCL20 mediates RANK/RANKL-induced epithelial-mesenchymal transition in endometrial cancer cells Oncotarget, 7(18):25328-39.
PMID: 27015366 -
Species: Human
Sample Types: Cell Culture Supernates
Chen X et al. (0), Melatonin attenuates hypoxia-induced epithelial-mesenchymal transition and cell aggressive via Smad7/ CCL20 in glioma. Oncotarge8(55):93580-93592.
PMID: 29212174
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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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Verified Customer | Posted 12/06/2021
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Verified Customer | Posted 03/25/2021
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Verified Customer | Posted 01/29/2021
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Verified Customer | Posted 04/20/2020This 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.
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Verified Customer | Posted 03/05/2020Gives very useful information. used with MDA-MB-231 cell lysate.
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Verified Customer | Posted 07/31/2018
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Verified Customer | Posted 03/08/2017
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FAQs for Proteome Profiler Human XL Oncology Array
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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.
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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.
