Proteome Profiler Human sReceptor Array, Hematopoietic
Proteome Profiler Human sReceptor Array, Hematopoietic Summary
To verify simultaneously detect the relative levels of 105 different soluble receptors expressed and released by hematopoietic cells in a single sample. No specialized equipment is necessary.
- Product Insert
- Troubleshooting Guide
- Common assays for analytes represented in the Human Common Analyte Array
- Assays for analytes represented in the Human sReceptor Array Kit Hematopoietic Panel
General Assay Principle
Carefully selected capture antibodies have been spotted in duplicate on nitrocellulose membranes. Cell culture supernatant, cell lysate, or serum samples are diluted and mixed with a cocktail of biotinylated detection antibodies. The sample/antibody mixture is then incubated with the array. Any cytokine/detection antibody complex present is bound by its cognate immobilized capture antibody on the membrane. Streptavidin-Horseradish Peroxidase and chemiluminescent detection reagents are added, and a signal is produced in proportion to the amount of cytokine bound. Chemiluminescence is detected in the same manner as a Western blot.
- 2 Hematopoeitic Array and Membranes
- 2 Common Analyte Array Membranes
- 4-Well Multi-dish
- Array Buffers
- Wash Buffer
- Detection Antibody Cocktail
- Streptavidin-HRP
- Chemiluminescent Detection Reagents
- Transparency Overlay Template
- Detailed Protocol
For a complete list of the kit contents and necessary materials, please see the Materials Provided and Other Materials Required sections of the product datasheet.
| Simultaneously detect the levels of these soluble receptors in a single sample. | |||||
| Hematopoeitic Array | Common Analytes Array | ||||
| CD5 | Integrin alpha 5/CD49e | ACE | HB-EGF | ||
| CD6 | Integrin alpha 6/CD49f | ADAM8 | ICAM-2/CD102 | ||
| CD30 | Integrin alpha 9 | ADAM9 | IL-1 RII | ||
| CD40 | Integrin alpha E/CD103 | ADAM10 | IL-15 R alpha | ||
| CD43 | Integrin alpha L/CD11a | ALCAM/CD166 | Integrin beta 1/CD29 | ||
| CD48/SLAMF2 | Integrin alpha M/CD11b | Amphiregulin | Integrin beta 2/CD18 | ||
| CD59 | Integrin alpha X/CD11c | APP (pan) | Integrin beta 3/CD61 | ||
| CD84/SLAMF5 | LAG-3 | BACE-1 | Integrin beta 4/CD104 | ||
| CD97 | MMP-9 (total) | BCAM/CD239 | Integrin beta 5 | ||
| CD163 | MMR | C1q R1/CD93 | Integrin beta 6 | ||
| CD229/SLAMF3 | Myeloperoxidase | CD9 | JAM-A/CD321 | ||
| Chitinase 3-like 1/YKL40 | Resistin | CD23/Fc epsilon RII | Lipocalin-2/NGAL | ||
| CRP | L-Selectin | CD31/PECAM-1 | LOX-1/SR-E1/CLEC8A | ||
| CRTAM | Siglec-5 | CD36/SR-B3 | MD-1/LY86 | ||
| CXCL16 | Siglec-6 | CD40 Ligand | MMP-2 (total) | ||
| DNAM-1 | Siglec-7 | CD44H | NCAM-1/CD56 | ||
| DPPIV/CD26 | Siglec-9 | CD58/LFA-3 | NCAM-L1/CD171 | ||
| IFN-gamma R2 | Siglec-10 | CD90/Thy1 | Osteopontin | ||
| IL-2 R alpha | TIM-3/KIM-3 | CD99 | PAR1 | ||
| IL-2 R beta | TLR2/CD282 | CD155/PVR | Pref-1/DLK-1/FA1 | ||
| IL-6 R | TLR4/CD284 | CEACAM-1/CD66a | RECK | ||
| Integrin alpha 3/CD49c | TNF RI | CX3CL1/Fractalkine | Stabilin-1 | ||
| Integrin alpha 4/CD49d | TRACP/PAP/ACP5 | CXCL8/IL-8 | TACE/ADAM17 | ||
| TRANCE/RANKL | EMMPRIN/CD147 | Thrombospondin-1 | |||
| TREM-1 | Endoglin/CD105 | TIMP-1 | |||
| Epiregulin | TIMP-2 | ||||
| Galectin-1 | TIMP-3 | ||||
| Galectin-3 | TNF RII | ||||
| Galectin-3BP/MAC-2BP | |||||
Specifications
Product Datasheets
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, relative expression levels of human soluble receptors 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
- 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.
Citations for Proteome Profiler Human sReceptor Array, Hematopoietic
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Stromal mesenchymal stem cells facilitate pancreatic cancer progression by regulating specific secretory molecules through mutual cellular interaction
Authors: K Saito, M Sakaguchi, S Maruyama, H Iioka, EW Putranto, IW Sumardika, N Tomonobu, T Kawasaki, K Homma, E Kondo
J Cancer, 2018-07-30;9(16):2916-2929. 2018-07-30
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Signaling via PI3K/FOXO1A pathway modulates formation and survival of human embryonic stem cell-derived endothelial cells.
Authors: Merkely B, Gara E, Lendvai Z, Skopal J, Leja T, Zhou W, Kosztin A, Varady G, Mioulane M, Bagyura Z, Nemeth T, Harding S, Foldes G
Stem Cells Dev, 2014-12-22;24(7):869-78. 2014-12-22
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