| Description | BCIP/NBT (5-bromo-4-chloro-3-indolyl phosphate/ Nitro Blue Tetrazolium) liquid ready-to-use substrate is a highly active and stable formulation utilized for colorimetric detection of Alkaline Phosphatase (AP) activity in membrane assays. Positive reactions form an intense blue/purple precipitate at BCIP/NBT (5-bromo-4-chloro-3-indolyl phosphate/ Nitro Blue Tetrazolium) liquid ready-to-use substrate is a highly active and stable formulation utilized for colorimetric detection of Alkaline Phosphatase (AP) activity in membrane assays. Positive reactions form an intense blue/purple precipitate at the site of the reaction. The color develops when AP catalyzes the dephosphorylation of BCIP and converts NBT to insoluble blue/purple NBT formazan. The intense blue/purple precipitate is very stable and resists fading when exposed to light.Product Characteristics BCIP/NBT (5-bromo-4-chloro-3-indolyl phosphate/ Nitro Blue Tetrazolium) liquid, ready-to-use substrate is a highly active and stable formulation utilized for colorimetric detection of Alkaline Phosphatase (AP) activity in membrane assays. Positive reactions form an intense blue/purple precipitate at the site of the reaction. The color develops when AP catalyzes the dephosphorylation of BCIP and converts NBT to insoluble blue/purple NBT formazan. The intense blue/purple precipitate is very stable and resists fading when exposed to light.Composition & Properties The BCIP/NBT is a liquid, ready-to-use substrate. The solution contains NBT, BCIP and non-toxic stabilizers in an aqueous buffer, pH 9.6. The formulation is free of organic solvents.Working Procedure 1.Allow the solution to reach room temperature prior to use. 2.After the final incubation with the AP-labelled probe, wash the membrane thoroughly in a Tris-Buffered Saline (TBS Buffer) containing 0.1% Tween 20. 3.After the final wash, completely cover the membrane with BCIP/NBT solution and allow the color reaction to develop until optimal signal intensity is reached (usually 5-30 minutes). Incubation time will depend on enzyme activity. A further adjustment of the dilution of the AP probe may be necessary. The formazan deposit may flake off the membrane or background staining of the membrane may occur if the color development is too fast or too slow, respectively. 4.Stop the enzyme reaction by thoroughly washing membrane with deionized water. 5.Dry the membrane and store at room temperature. It is recommended to protect the membrane from light for a permanent record of results.Tips & Tricks Normal appearance of the solution is clear, pale yellow. Discard if solution is turbid or purple. Crystalline precipitate can occur in the product. Do not use phosphate buffers, as inorganic phosphate is a powerful inhibitor of AP. The products are not recommended for immunohistochemical or in situ hybridization techniques.Handling & Storage Store solution at 2-8 °C in the dark. Avoid exposure to light and heat. Re-dispense only into bottles made of High Density Polyethylene (HDPE), amber color. Dispensing guidelines are available upon request... Read More | Inquire | Inquire | Purity>95% (SDS-PAGE&HPLC) Endotoxin level<0.1 EU/µgFunctionMay regulate apoptosis, cell proliferation and cell differentiation. Binds beta-galactoside and a wide array of complex carbohydrates. Inhibits CD45 protein phosphatase activity and therefore the dephosphorylation of Lyn Purity>95% (SDS-PAGE&HPLC) Endotoxin level<0.1 EU/µgFunctionMay regulate apoptosis, cell proliferation and cell differentiation. Binds beta-galactoside and a wide array of complex carbohydrates. Inhibits CD45 protein phosphatase activity and therefore the dephosphorylation of Lyn kinase.Gal-1 is also engaged in many protein-protein interactions. Gal-1 plays a number of crucial roles in neuronal cell differentiation and survival in both the central and the peripheral nervous systems, and the establishment and maintenance of T-cell tolerance and homeostasis in vivo... Read More | BackgroundStreptavidin is a tetrameric bacterial protein isolated from Streptomyces avidinii providing 4 high-affinity biotin binding sites. Streptavidin homo-tetramers have an extraordinarily high affinity for biotin. With a dissociation constant on the order of ≈10⁻¹⁴ mol/L,BackgroundStreptavidin is a tetrameric bacterial protein isolated from Streptomyces avidinii providing 4 high-affinity biotin binding sites. Streptavidin homo-tetramers have an extraordinarily high affinity for biotin. With a dissociation constant on the order of ≈10⁻¹⁴ mol/L, the binding of biotin to streptavidin is one of the strongest non-covalent interactions known in nature. Unlike egg-white avidin, which has a net positive charge at neutral pH and contains about 7% carbohydrate, streptavidin has almost no net charge at neutral pH, does not contain carbohydrate, and exhibits lower non-specific background. Streptavidin conjugates are widely used together with a conjugate of biotin for specific detection of a variety of proteins, protein motifs, nucleic acids and other molecules. This FITC-streptavidin conjugate was prepared by highly purified Streptavidin and free FITC was removed. Streptavidin (FITC) is a useful second-step reagent for the indirect immunofluorescent staining of cells in combination with biotinylated primary antibodies for flow cytometric analysis. Excitation at 488nm light leads to a fluorescence emission maximum of 520 nm.Recommended Usage：Every lot of Streptavidin-FITC is tested by flow cytometry using biotinylated primary antibodies. From this testing it is recommended that between 0.02 and 0.25 µg of streptavidin be used per 106 cells in a 100 µl staining volume... Read More |