Keep your chicken LDH samples on ice during the experiment. However, the Bradford assays are performed...

  

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Keep your chicken LDH samples on ice during the experiment. However, the Bradford assays are performed with reagents that are at room temperature. Add your cold samples to the room temperature reagents in the assay tubes. 1. Retrieve the fractions labeled C.LDH Crude 3D B, CLDH-P-B, and C.LDH Purified 3D B and samples NADH 2 and NADH 3 stored in the 4 °C cold-room. Keep them on ice. 2. Calculate the amount of 200 µg/ml BSA Stock solution and 10 mM Tris-HCI buffer to add to each reaction shown in Table 1. Write this data into the table. 3. DO NOT add Bradford Reagent until ready for it! Pipette the volumes of BSA Stock and 10 mM Tris-HCl, pH 8.6 required into the test tubes provided, as shown in Table 1. Table 1. Information for construction of a standard curve. You must do the calculations for the amount of BSA stock and Tris-HCI buffer to add to each assay. Assay # Protein 200 Sample 10 mM Bradford A595 Added Type Tris- (µg) HCI, pH 8.6 123 +16 1 2 4 5 6 o al 0 2 4 630 8 10 µg/mL BSA Stock (μl) 0 Blank BSA BSA BSA BSA BSA 800 Reagent (μl) Add later 200 200 200 200 200 200 Final Volum e (mL) 1.0 1.0 1.0 1.0 1.0 1.0 Keep your chicken LDH samples on ice during the experiment. However, the Bradford assays are performed with reagents that are at room temperature. Add your cold samples to the room temperature reagents in the assay tubes. 1. Retrieve the fractions labeled C.LDH Crude 3D B, CLDH-P-B, and C.LDH Purified 3D B and samples NADH 2 and NADH 3 stored in the 4 °C cold-room. Keep them on ice. 2. Calculate the amount of 200 µg/ml BSA Stock solution and 10 mM Tris-HCI buffer to add to each reaction shown in Table 1. Write this data into the table. 3. DO NOT add Bradford Reagent until ready for it! Pipette the volumes of BSA Stock and 10 mM Tris-HCl, pH 8.6 required into the test tubes provided, as shown in Table 1. Table 1. Information for construction of a standard curve. You must do the calculations for the amount of BSA stock and Tris-HCI buffer to add to each assay. Assay # Protein 200 Sample 10 mM Bradford A595 Added Type Tris- (µg) HCI, pH 8.6 123 +16 1 2 4 5 6 o al 0 2 4 630 8 10 µg/mL BSA Stock (μl) 0 Blank BSA BSA BSA BSA BSA 800 Reagent (μl) Add later 200 200 200 200 200 200 Final Volum e (mL) 1.0 1.0 1.0 1.0 1.0 1.0