Proteome Profiler Mouse XL Cytokine Array
Proteome Profiler Mouse XL Cytokine Array Summary
A membrane-based antibody array for the parallel determination of the relative levels of selected mouse cytokines and chemokines. Validated for analyte detection in cell culture supernates, cell lysates, tissue lysates, serum, and urine.
Troubleshooting GuideKey Benefits
- Detects 111 mouse cytokines simultaneously
- Requires no specialized equipment
- Compatible with LI-COR* and chemiluminescence detection
Principle of the Assay
The Proteome Profiler Mouse XL Cytokine Array Kit is a membrane-based sandwich immunoassay. Capture antibodies spotted in duplicate on nitrocellulose membranes bind to specific target proteins present in the sample (Step 1). Captured proteins are detected with biotinylated detection antibodies (Step 2) and then visualized using chemiluminescent detection reagents (Step 3). The signal produced is proportional to the amount of analyte bound.
Why Use an Antibody Array to Detect Multiple Cytokines?
Determining the expression of multiple cytokines in a single sample can be expensive, time consuming and can require specialized equipment. Performing multiple immunoprecipitations and Western blots requires time, labor, and reagents. The use of a multiplex antibody array to detect multiple cytokines in a single sample can be cost-effective and also save time and sample.
- 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)
For a complete list of the kit contents and necessary materials, please see the Materials Provided/Other Supplies Required sections of the product datasheet.
Simultaneously detect the levels of these cytokines, chemokines, and growth factors in a single sample. | ||
Adiponectin/Acrp30 | DPPIV/CD26 | IL-27 |
Amphiregulin | EGF | IL-28 |
Angiopoietin-1 | Endoglin/CD105 | IL-33 |
Angiopoietin-2 | Endostatin | LDL R |
Angiopoietin-like 3 | Fetuin A/AHSG | Leptin |
BAFF/BLyS/TNFSF13B | FGF acidic | LIF |
C1q R1/CD93 | FGF-21 | Lipocalin-2/NGAL, |
CCL2/JE/MCP-1 | Flt-3 Ligand | LIX |
CCL3/CCL4 MIP-1 alpha/beta | Gas6 | M-CSF |
CCL5/RANTES | G-CSF | MMP-2 |
CCL6/C10 | GDF-15 | MMP-3 |
CCL11/Eotaxin | GM-CSF | MMP-9 |
CCL12/MCP-5 | HGF | Myeloperoxidase |
CCL17/TARC | ICAM-1/CD54 | Osteopontin (OPN) |
CCL19/MIP-3 beta | IFN-gamma | Osteoprotegerin/TNFRSF11B |
CCL20/MIP-3 alpha | IGFBP-1 | PD-ECGF/Thymidine phosphorylase |
CCL21/6Ckine | IGFBP-2 | PDGF-BB |
CCL22/MDC | IGFBP-3 | Pentraxin 2/SAP |
CD14 | IGFBP-5 | Pentraxin 3/ TSG-14 |
CD40/TNFRSF5 | IGFBP-6 | Periostin/OSF-2 |
CD160 | IL-1 alpha/IL1F1 | Pref-1/DLK-1/FA1 |
Chemerin | IL-1 beta/IL-1F2 | Proliferin |
Chitinase 3-like 1 | IL-1ra/IL-1F3 | Proprotein Convertase 9/PCSK9 |
Coagulation Factor III/Tissue Factor | IL-2 | RAGE |
Complement Component C5/C5a | IL-3 | RBP4 |
Complement Factor D | IL-4 | Reg3G |
C-Reactive Protein/CRP | IL-5 | Resistin |
CX3CL1/Fractalkine | IL-6 | E-Selectin/CD62E |
CXCL1/KC | IL-7 | P-Selectin/CD62P |
CXCL2/MIP-2 | IL-10 | Serpin E1/PAI-1 |
CXCL9/MIG | IL-11 | Serpin F1/PEDF |
CXCL10/IP-10 | IL-12p40 | Thrombopoietin |
CXCL11/I-TAC | IL-13 | TIM-1/KIM-1/HAVCR |
CXCL13/BLC/BCA-1 | IL-15 | TNF-alpha |
CXCL16 | IL-17A | VCAM-1/CD106 |
Cystatin C | IL-22 | VEGF |
Dkk-1 | IL-23 | WISP-1/CCN4 |
Assays for Analytes represented in the Mouse XL Cytokine Array Kit
Analyte | Qkit | Duoset | ||
Adiponectin/Acrp30 | MRP300 | DY1119 | ||
Amphiregulin | DY989 | |||
Angiopoietin-1 | ||||
Angiopoietin-2 | ||||
Angiopoietin-like 3 | MANL30 | DY136 | ||
BAFF/BLyS/TNFSF13B | MBLYS0 | DY2106-05 | ||
C1q R1/CD93 | MCD930 | |||
CCL2/JE/MCP-1 | MJE00 | DY479 | ||
CCL3/CCL4 MIP-1 alpha/beta | ||||
CCL5/RANTES | MMR00 | DY478 | ||
CCL6/C10 | DY487 | |||
CCL11/Eotaxin | MME00 | DY420 | ||
CCL12/MCP-5 | MCC120 | DY428 | ||
CCL17/TARC | MCC170 | DY529 | ||
CCL19/MIP-3 beta | DY440 | |||
CCL20/MIP-3 alpha | MCC200 | DY760 | ||
CCL21/6Ckine | DY457 | |||
CCL22/MDC | MCC220 | DY439 | ||
CD14 | MC140 | DY982 | ||
CD40/TNFRSF5 | MCCD40 | DY097 | ||
CD160 | MCHM00 | DY2325 | ||
Chemerin | ||||
Chitinase 3-like 1 | MC3L10 | DY2649 | ||
Coagulation Factor III/Tissue Factor | ||||
Complement Component C5/C5a | ||||
Complement Factor D | ||||
C-Reactive Protein/CRP | MCRP00 | DY1829 | ||
CX3CL1/Fractalkine | MCX310 | DY472 | ||
CXCL1/KC | MKC00B | DY453 | ||
CXCL2/MIP-2 | MM200 | DY452 | ||
CXCL9/MIG | MCX900 | DY492 | ||
CXCL10/IP-10 | MCX100 | DY466 | ||
CXCL11/I-TAC | DY572 | |||
CXCL13/BLC/BCA-1 | MCX130 | |||
CXCL16 | DY503 | |||
Cystatin C | MSCTC0 | DY1238 | ||
Dkk-1 | MKK100 | DY1765 | ||
DPPIV/CD26 | DY954 | |||
EGF | MEG00 | DY2028 | ||
Endoglin/CD105 | MNDG00 | DY1320 | ||
Endostatin | ||||
Fetuin A/AHSG | MFTA00 | DY1563 | ||
FGF acidic | DY4686-05 | |||
FGF-21 | MF2100 | DY3057 | ||
Flt-3 Ligand | MFK00 | DY427 | ||
Gas6 | MGAS60 | DY986 | ||
G-CSF | MCS00 | DY414 | ||
GDF-15 | MGD150 | DY6385 | ||
GM-CSF | MGM00 | DY415 | ||
HGF | MHG00 | DY2207 | ||
ICAM-1/CD54 | MIC100 | DY796 | ||
IFN-gamma | MIF00 | DY485 | ||
IGFBP-1 | DY1588-05 | |||
IGFBP-2 | DY797 | |||
IGFBP-3 | MGB300 | DY775 | ||
IGFBP-5 | DY578 | |||
IGFBP-6 | DY776 | |||
IL-1 alpha/IL1F1 | MLA00 | DY400 | ||
IL-1 beta/IL-1F2 | MLB00C | DY401 | ||
IL-1ra/IL-1F3 | MRA00 | DY480 | ||
IL-2 | M2000 | DY402 | ||
IL-3 | M3000 | DY403 | ||
IL-4 | M4000B | DY404 | ||
IL-5 | M5000 | DY405 | ||
IL-6 | M6000B | DY406 | ||
IL-7 | M7000 | DY407 | ||
IL-10 | M1000B | DY417 | ||
IL-11 | DY418 | |||
IL-12p40 | MP400 non-allele specific |
M1240 allele specific |
DY2398 non-allele specific |
DY499 allele specific |
IL-13 | M1300CB | DY413 | ||
IL-15 | DY447 | |||
IL-17A | M1700 | DY421 | ||
IL-22 | M2200 | DY582 | ||
IL-23 | M2300 | DY1887 | ||
IL-27 | DY1789B | |||
IL-28 | ||||
IL-33 | M3300 | DY3626 | ||
LDL R | MLDLR0 | DY2255 | ||
Leptin | MOB00 | DY498 | ||
LIF | MLF00 | |||
Lipocalin-2/NGAL, | DY1857 | |||
LIX | MX000 | DY443 | ||
M-CSF | MMC00 | DY416 | ||
MMP-2 | MMP200 | |||
MMP-3 | MMP300 | DY548 | ||
MMP-9 | MMPT90 | MMP900B | DY6718 | DY909 |
Myeloperoxidase | DY3667 | |||
Osteopontin (OPN) | MOST00 | DY441 | ||
Osteoprotegerin/TNFRSF11B | MOP00 | DY459 | ||
PD-ECGF/Thymidine phosphorylase | ||||
PDGF-BB | MBB00 | |||
Pentraxin 2/SAP | MPTX20 | |||
Pentraxin 3/ TSG-14 | MPTX30 | DY2166 | ||
Periostin/OSF-2 | MOSF20 | DY2955 | ||
Pref-1/DLK-1/FA1 | ||||
Proliferin | ||||
Proprotein Convertase 9/PCSK9 | MPC900 | DY3985 | ||
RAGE | MRG00 | DY1179 | ||
RBP4 | MRBP40 | DY3476 | ||
Reg3G | ||||
Resistin | MRSN00 | DY1069 | ||
E-Selectin/CD62E | MES00 | DY575 | ||
P-Selectin/CD62P | MPS00 | DY737 | ||
Serpin E1/PAI-1 | DY3828-05 | |||
Serpin F1/PEDF | ||||
Thrombopoietin | MTP00 | DY488 | ||
TIM-1/KIM-1/HAVCR | MKM100 | DY1817 | ||
TNF-alpha | MTA00B | DY410 | ||
VCAM-1/CD106 | MVC00 | DY643 | ||
VEGF | MMV00 | DY493 | ||
WISP-1/CCN4 | MWSP10 |
Specifications
Product Datasheets
Scientific Data
Detection of Selected Mouse Cytokines in Balb/3T3 Cell Line Lysate. Lysates from Balb/3T3 mouse embryonic fibroblast cells were untreated or treated with 100 ng/mL of recombinant mouse TNF-alpha (R&D Systems, Catalog # 410-MT) for 24 hours (200 µg lysate, 10 minute exposure).
Detection of Selected Mouse Cytokines in CMT-93 Cell Line Lysate. Lysates from CMT-93 mouse rectal carcinoma cells were untreated or treated with 100 ng/mL LPS for 24 hours (200 µg lysate, 10 minute exposure).
Detection of Selected Mouse Cytokines in Balb/3T3 Cell Line Supernate. Balb/3T3 mouse embryonic fibroblast cells were untreated or treated with 100 ng/mL of recombinant mouse TNF-alpha (R&D Systems, Catalog # 410-MT) for 24 hours (500 µL of cell culture supernate, 5 minute exposure).
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
- 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)
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
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.
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.
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 Mouse XL Cytokine Array
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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The Effect of Super-Repressor IkB-Loaded Exosomes (Exo-srIkappaBs) in Chronic Post-Ischemia Pain (CPIP) Models
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Microglia and macrophages promote corralling, wound compaction and recovery after spinal cord injury via Plexin-B2
Authors: X Zhou, S Wahane, MS Friedl, M Kluge, CC Friedel, K Avrampou, V Zachariou, L Guo, B Zhang, X He, RH Friedel, H Zou
Nat. Neurosci., 2020-03-01;23(3):337-350. 2020-03-01
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beta4GALT1 controls beta1 integrin function to govern thrombopoiesis and hematopoietic stem cell homeostasis
Authors: S Giannini, MM Lee-Sundlo, L Rivadeneyr, CA Di Buduo, R Burns, JT Lau, H Falet, A Balduini, KM Hoffmeiste
Nat Commun, 2020-01-17;11(1):356. 2020-01-17
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beta4GALT1 controls beta1 integrin function to govern thrombopoiesis and hematopoietic stem cell homeostasis
Authors: S Giannini, MM Lee-Sundlo, L Rivadeneyr, CA Di Buduo, R Burns, JT Lau, H Falet, A Balduini, KM Hoffmeiste
Nat Commun, 2020;11(1):356. 2020
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Dual microglia effects on blood brain barrier permeability induced by systemic inflammation
Authors: K Haruwaka, A Ikegami, Y Tachibana, N Ohno, H Konishi, A Hashimoto, M Matsumoto, D Kato, R Ono, H Kiyama, AJ Moorhouse, J Nabekura, H Wake
Nat Commun, 2019-12-20;10(1):5816. 2019-12-20
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Depletion of Bone Marrow-Derived Fibrocytes Attenuates TAA-Induced Liver Fibrosis in Mice
Authors: F Hempel, M Roderfeld, R Savai, A Sydykov, K Irungbam, R Schermuly, R Voswinckel, K Köhler, Y Churin, L Kiss, J Bier, J Pons-Kühne, E Roeb
Cells, 2019-10-07;8(10):. 2019-10-07
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Non-classical tissue monocytes and two functionally distinct populations of interstitial macrophages populate the mouse lung
Authors: J Schyns, Q Bai, C Ruscitti, C Radermecke, S De Scheppe, S Chakarov, F Farnir, D Pirottin, F Ginhoux, G Boeckxstae, F Bureau, T Marichal
Nat Commun, 2019-09-03;10(1):3964. 2019-09-03
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Acidity promotes tumour progression by altering macrophage phenotype in prostate cancer
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Br. J. Cancer, 2019-08-16;121(7):556-566. 2019-08-16
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Tumour-Derived Laminin alpha5 (LAMA5) Promotes Colorectal Liver Metastasis Growth, Branching Angiogenesis and Notch Pathway Inhibition
Authors: A Gordon-Wee, SY Lim, A Yuzhalin, S Lucotti, JAF Vermeer, K Jones, J Chen, RJ Muschel
Cancers (Basel), 2019-05-06;11(5):. 2019-05-06
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Platelet GPIb? is a mediator and potential interventional target for NASH and subsequent liver cancer
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Nat. Med., 2019-04-01;25(4):641-655. 2019-04-01
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Liver-derived fibroblast growth factor 21 mediates effects of glucagon-like peptide-1 in attenuating hepatic glucose output
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EBioMedicine, 2019-03-01;0(0):. 2019-03-01
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Prostate cancer promotes a vicious cycle of bone metastasis progression through inducing osteocytes to secrete GDF15 that stimulates prostate cancer growth and invasion
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Oncogene, 2019-02-12;0(0):. 2019-02-12
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Matrix bound nanovesicle-associated IL-33 activates a pro-remodeling macrophage phenotype via a non-canonical, ST2-independent pathway
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J Immunol Regen Med, 2019-02-01;3(0):26-35. 2019-02-01
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T cell microvilli constitute immunological synaptosomes that carry messages to antigen-presenting cells
Authors: HR Kim, Y Mun, KS Lee, YJ Park, JS Park, JH Park, BN Jeon, CH Kim, Y Jun, YM Hyun, M Kim, SM Lee, CS Park, SH Im, CD Jun
Nat Commun, 2018-09-07;9(1):3630. 2018-09-07
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IL-23 secreted by myeloid cells drives castration-resistant prostate cancer
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Nature, 2018-06-27;0(0):. 2018-06-27
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Extracellular vesicles mediate low dose ionizing radiation-induced immune and inflammatory responses in the blood
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Int. J. Radiat. Biol., 2018-03-29;0(0):1-31. 2018-03-29
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Comparison of the immune response during acute and chronic Staphylococcus aureus infection
Authors: RA Brady, CP Mocca, RD Plaut, K Takeda, DL Burns
PLoS ONE, 2018-03-29;13(3):e0195342. 2018-03-29
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Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis
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Front Immunol, 2018-03-06;9(0):334. 2018-03-06
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Mesenchymal stem cells over-expressing cxcl12 enhance the radioresistance of the small intestine
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Cell Death Dis, 2018-02-05;9(2):154. 2018-02-05
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Conditioned Medium of Bone Marrow-Derived Mesenchymal Stromal Cells as a Therapeutic Approach to Neuropathic Pain: A Preclinical Evaluation
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Stem Cells Int, 2018-01-30;2018(0):8179013. 2018-01-30
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Tumor-extrinsic discoidin domain receptor 1 promotes mammary tumor growth by regulatingadipose stromal interleukin 6 production in mice
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A CREB3-regulated ER-Golgi trafficking signature promotes metastatic progression in breast cancer
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Pharmacological HIF-inhibition attenuates postoperative adhesion formation
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A Jak1/2 inhibitor, baricitinib, inhibits osteoclastogenesis by suppressing RANKL expression in osteoblasts in vitro
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Map3k8 controls granulocyte colony-stimulating factor production and neutrophil precursor proliferation in lipopolysaccharide-induced emergency granulopoiesis
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Promyelocytic Leukemia Protein (PML) Controls Listeria monocytogenes Infection
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MBio, 2017-01-10;8(1):. 2017-01-10
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Depletion of microglia exacerbates postischemic inflammation and brain injury
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Astrocyte-derived interleukin-15 exacerbates ischemic brain injury via propagation of cellular immunity
Proc. Natl. Acad. Sci. U.S.A, 2016-12-19;0(0):. 2016-12-19
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Novel Resolvin D2 Receptor Axis in Infectious Inflammation
Authors: Nan Chiang
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Evaluation of PLGA containing anti-CTLA4 inhibited endometriosis progression by regulating CD4+CD25+Treg cells in peritoneal fluid of mouse endometriosis model
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FAQs
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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.
Reviews for Proteome Profiler Mouse XL Cytokine Array
Average Rating: 4.9 (Based on 35 Reviews)
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Performed on mammary tumor lysates.
I bought two boxes, and they're both great
This product works very well
Conditioned medium of macrophages (36h) isolated from metastatic lungs.
This kit is really great!
Secretome of 4T1 murine breast cancer cells.
Cytokine array of the secretome of MLE12 cells.
We used the array kit for profiling inflammatory cytokines in plasma samples from mice. I trust that it works very well.
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.
Excellent specificity and detection for large number of cytokines. Works perfectly with little optimization.
Pancreas lysate. 750ug protein/ membrane
200 uL pooled serum from wild-type C57Bl/6 mice.
5 min exposure time
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.