Purpose | C-Series Human Matrix Metalloproteinase Antibody Array 1 Kit. Detects 10 Human MMPs and TIMPs. Suitable for all liquid sample types. |
Brand | RayBio? |
Sample Type | Serum, Plasma, Cell Culture Supernatant, Cell Lysate, Tissue Lysate |
Analytical Method | Semi-Quantitative |
Detection Method | Chemiluminescent |
Specificity | MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-10, MMP-13, TIMP-1, TIMP-2, TIMP-4 |
Characteristics |
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Components |
Antibody Array Membranes Biotinylated Detection Antibody Cocktail Blocking Buffer Wash Buffers 1 and 2 Cell & Tissue Lysis Buffer Detection Buffers C and D Plastic Incubation Tray Protease Inhibitor Cocktail (in select kits) |
Material not included |
Pipettors, pipet tips and other common lab consumables Orbital shaker or oscillating rocker Tissue Paper, blotting paper or chromatography paper Adhesive tape or Saran Wrap Distilled or de-ionized water A chemiluminescent blot documentation system (such as UVP's ChemiDoc-It? or EpiChem II Benchtop Darkroom), X-ray Film and a suitable film processor, or another chemiluminescent detection system. |
Application Notes |
Perform ALL incubation and wash steps under gentle rotation or rocking motion (~0.5 to 1 cycle/sec) using an orbital shaker or oscillating rocker to ensure complete and even reagent/sample coverage. Rocking/rotating too vigorously may cause foaming or bubbles to appear on the membrane surface which, should be avoided. All washes and incubations should be performed in the Incubation Tray (ITEM 10) provided in the kit. Cover the Incubation Tray with the lid provided during all incubation steps to avoid evaporation and outside debris contamination. Ensure the membranes are completely covered with sufficient sample or reagent volume during each incubation. Avoid forceful pipetting directly onto the membrane, instead, gently pipette samples and reagents into a corner of each well. Aspirate samples and reagents completely after each step by suctioning off excess liquid with a pipette. Tilting the tray so the liquid moves to a corner and then pipetting is an effective method. Optional overnight incubations may be performed for the following step to increase overall spot signal intensities: - Sample Incubation - Biotinylated Antibody Cocktail Incubation - HRP-Streptavidin Incubation |
Comment |
The C-Series arrays feature chemiluminescent signal detection. The antibodies are spotted on nitrocellulose membrane solid supports and are handled in a very similar manner to Western blots. |
Sample Volume | 1 mL |
Plate | Membrane |
Protocol |
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Sample Preparation |
Use serum-free conditioned media if possible. If serum-containing conditioned media is required, it is highly recommended that complete medium be used as a control since many types of sera contains cytokines. We recommend the following parameters for your samples: 50 to 100 μl of original or diluted serum, plasma, cell culture media, or other body fluid, or 50-500 μg/ml of protein for cell and tissue lysates. If you experience high background or if the fluorescent signal intensities exceed the detection range, further dilution of your sample is recommended. |
Assay Procedure |
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Calculation of Results |
Visual comparison of array images may be sufficient to see differences in relative protein expression. However, most researchers will want to perform numerical comparisons of the signal intensities (or more precisely, signal densities), using 2-D densitometry. Gel/Blot documentation systems and other chemiluminescent or phosphorescent detection systems are usually sold as a package with compatible densitometry software. Any densitometry software should be sufficient to obtain spot signal densities from your scanned images. One such software program, ImageJ, is available for free from the NIH website along with an array plug-in. |
Assay Precision | Inter-array Coefficient of Variation (CV) of spot signal intensities as low as 5% when run under optimal conditions. |
Restrictions | For Research Use only |
Handling Advice | The antibody printed side of each membrane is marked by a dash (-) or number (#) in the upper left corner. Do not allow membranes to dry out during the experiment or they may become fragile and break OR high and/or uneven background may occur. Grasp membranes by the corners or edges only using forceps. DO NOT touch printed antibody spots. |
Storage | -20 °C |
Storage Comment | For best results, store the entire kit frozen at -20°C upon arrival. Stored frozen, the kit will be stable for at least 6 months which is the duration of the product warranty period. Once thawed, store array membranes and 1X Blocking Buffer at -20°C and all other reagents undiluted at 4°C for no more than 3 months. |
Expiry Date | 6 months |
Supplier Images |
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Product cited in: |
Salhi, Farhadian, Giles, Vega-Saenz de Miera, Silva, Bourque, Yeh, Chhangawala, Wang, Ye, Zhang, Hernando-Monge, Houvras, Osman: "RSK1 activation promotes invasion in nodular melanoma." in: The American journal of pathology, Vol. 185, Issue 3, pp. 704-16, 2015 (PubMed).
Perrigue, Silva, Warden, Feng, Reid, Mota, Joseph, Tian, Glackin, Gutova, Najbauer, Aboody, Barish: "The Histone Demethylase Jumonji Coordinates Cellular Senescence Including Secretion of Neural Stem Cell-attracting Cytokines." in: Molecular cancer research : MCR, 2015 (PubMed). Huang, Yang, Xiang, Peng, Qiu, Wan, Zhang, Li, Li, Ren: "Diallyl disulfide inhibits growth and metastatic potential of human triple-negative breast cancer cells through inactivation of the ?-catenin signaling pathway." in: Molecular nutrition & food research, Vol. 59, Issue 6, pp. 1063-75, 2015 (PubMed). Yang, Huang, Xiang, Yin, Luo, Huang, Luo, Li, Li, Ren: "Chrysin inhibits metastatic potential of human triple-negative breast cancer cells by modulating matrix metalloproteinase-10, epithelial to mesenchymal transition, and PI3K/Akt signaling pathway." in: Journal of applied toxicology : JAT, Vol. 34, Issue 1, pp. 105-12, 2014 (PubMed). Carduner, Leroy-Dudal, Picot, Gallet, Carreiras, Kellouche: "Ascites-induced shift along epithelial-mesenchymal spectrum in ovarian cancer cells: enhancement of their invasive behavior partly dependant on αv integrins." in: Clinical & experimental metastasis, Vol. 31, Issue 6, pp. 675-88, 2014 (PubMed). Sakimoto, Ohnishi, Ishimori: "Simultaneous study of matrix metalloproteinases, proinflammatory cytokines, and soluble cytokine receptors in the tears of noninfectious corneal ulcer patients." in: Graefe's archive for clinical and experimental ophthalmology, Vol. 252, Issue 9, pp. 1451-6, 2014 (PubMed). Ichikawa, Sugiura, Koarai, Minakata, Kikuchi, Morishita, Oka, Kanai, Kawabata, Hiramatsu, Akamatsu, Hirano, Nakanishi, Matsunaga, Yamamoto, Ichinose: "TLR3 activation augments matrix metalloproteinase production through reactive nitrogen species generation in human lung fibroblasts." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 192, Issue 11, pp. 4977-88, 2014 (PubMed). Patil, Joshi, Kuna, Xu, Johannesson, Olausson, Sumitran-Holgersson: "CD271 identifies functional human hepatic stellate cells, which localize in peri-sinusoidal and portal areas in liver after partial hepatectomy." in: Cytotherapy, Vol. 16, Issue 7, pp. 990-9, 2014 (PubMed). De Boeck, Hendrix, Maynard, Van Bockstal, Dani?ls, Pauwels, Gespach, Bracke, De Wever: "Differential secretome analysis of cancer-associated fibroblasts and bone marrow-derived precursors to identify microenvironmental regulators of colon cancer progression." in: Proteomics, Vol. 13, Issue 2, pp. 379-88, 2013 (PubMed). Zhu, Hong, Tripathi, Sehdev, Belkhiri, El-Rifai: "Regulation of CXCR4-mediated invasion by DARPP-32 in gastric cancer cells." in: Molecular cancer research : MCR, Vol. 11, Issue 1, pp. 86-94, 2013 (PubMed). Vesey, Suen, Seow, Lohman, Liu, Gobe, Johnson, Fairlie: "PAR2-induced inflammatory responses in human kidney tubular epithelial cells." in: American journal of physiology. Renal physiology, Vol. 304, Issue 6, pp. F737-50, 2013 (PubMed). Lisboa, Andrade, Cunha-Melo: "Toll-like receptor activation and mechanical force stimulation promote the secretion of matrix metalloproteinases 1, 3 and 10 of human periodontal fibroblasts via p38, JNK and NF-kB." in: Archives of oral biology, Vol. 58, Issue 6, pp. 731-9, 2013 (PubMed). Mercuri, Patnaik, Dion, Gill, Liao, Simionescu: "Regenerative potential of decellularized porcine nucleus pulposus hydrogel scaffolds: stem cell differentiation, matrix remodeling, and biocompatibility studies." in: Tissue engineering. Part A, Vol. 19, Issue 7-8, pp. 952-66, 2013 (PubMed). Connolly, Mullan, McCormick, Matthews, Sullivan, Kennedy, FitzGerald, Poole, Bresnihan, Veale, Fearon: "Acute-phase serum amyloid A regulates tumor necrosis factor ? and matrix turnover and predicts disease progression in patients with inflammatory arthritis before and after biologic therapy." in: Arthritis and rheumatism, Vol. 64, Issue 4, pp. 1035-45, 2012 (PubMed). Caino, Lopez-Haber, Kissil, Kazanietz: "Non-small cell lung carcinoma cell motility, rac activation and metastatic dissemination are mediated by protein kinase C epsilon." in: PLoS ONE, Vol. 7, Issue 2, pp. e31714, 2012 (PubMed). Li, Ackerman, Mihai, Volakis, Ghadiali, Kniss: "Myoferlin depletion in breast cancer cells promotes mesenchymal to epithelial shape change and stalls invasion." in: PLoS ONE, Vol. 7, Issue 6, pp. e39766, 2012 (PubMed). Abdul Muneer, Alikunju, Szlachetka, Haorah: "The mechanisms of cerebral vascular dysfunction and neuroinflammation by MMP-mediated degradation of VEGFR-2 in alcohol ingestion." in: Arteriosclerosis, thrombosis, and vascular biology, Vol. 32, Issue 5, pp. 1167-77, 2012 (PubMed). Harper, Godwin, Green, Wilkes, Holden, Moffatt, Cookson, Layton, Chandler: "A study of matrix metalloproteinase expression and activity in atopic dermatitis using a novel skin wash sampling assay for functional biomarker analysis." in: The British journal of dermatology, Vol. 162, Issue 2, pp. 397-403, 2010 (PubMed). Raghu, Lakka, Gondi, Mohanam, Dinh, Gujrati, Rao: "Suppression of uPA and uPAR attenuates angiogenin mediated angiogenesis in endothelial and glioblastoma cell lines." in: PLoS ONE, Vol. 5, Issue 8, pp. e12458, 2010 (PubMed). Long, Blake, Song, Lark, Loeser: "Human articular chondrocytes produce IL-7 and respond to IL-7 with increased production of matrix metalloproteinase-13." in: Arthritis research & therapy, Vol. 10, Issue 1, pp. R23, 2008 (PubMed). |