Osteopontin (2AR, Bone Sialoprotein 1, Calcium Oxalate Crystal Growth Inhibitor Protein, Early T-lymphocyte Activation 1 Protein, Minopontin, Secreted Phosphoprotein 1, SPP-1, Spp1, Eta-1, Op, Spp-1) (PE)

Osteopontin (OPN), is a secreted acidic phosphorylated glycoprotein originally isolated from bone matrix. It has been shown to bind to different cell types through interactions with various integrins and CD44 variants. It has important functions in bone metabolism and inflammatory processes.\n\nApplications:\nSuitable for use in Flow Cytometry. Other applications not tested.\n\nRecommended Dilution:\nFlow Cytometry: 10ul labels 5x10e5 cells. Validated on RAW264.7 cells. For intracellular staining, cells must first be fixed and permeabilized. The use of 4% PFA as a fixative and a 0.1% saponin balanced salt solution for permeabilization and washing is recommended. \nOptimal dilutions to be determined by the researcher.\n\nFlow Cytometry Protocol:\n1. Cells were harvested and washed twice in saline buffer. \n2. Cell surface staining may be done at this point following the manufacturer's staining procedure. \n3. 5x10e5 cells were resuspended in 500ul of cold 4% paraformaldehyde fixation buffer (PFA) and incubated at RT for 10 minutes. \n4. Following fixation, cells were washed twice in saline buffer, then once in 0.1% saponin balanced salt solution. \n5. After permeabilization, 10ul of O8064-01N was added and the cells were incubated for 30 minutes at RT in the dark. \n6. The cells were washed twice with 0.1% saponin saline solution.\n7. The cells were resuspended in saline buffer for final flow cytometric analysis. As a control for analysis, cells in a separate tube should be treated with PE-labeled goat IgG antibody. \n\nStorage and Stability:\nMay be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Stable for at least 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.

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