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MCB120L exp 2 - 1 Standard Curves of UV and Bradford Assays...

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1) Standard Curves of UV and Bradford Assays: y = 0.656x - 0.001 R² = 0.998 -0.2 0 0.2 0.4 0.6 0.8 1 1.2 0 0.5 1 1.5 2 A b s280(co rec ted ) A280 Assay Series1 Linear (Series1) BSAmg/ml y = -0.010x 2 + 0.145x + 0.034 R² = 0.983 0 0.1 0.2 0.3 0.4 0.5 0.6 0 2 4 6 8 10 A b s595nm(corec ted ) BSA, ug Bradford Assay Series1 Poly. (Series1) 2) Concentration of original unknown protein: A280 Assay: 0.54 with Standard Deviation of 0.029 Bradford Assay: 0.66 with Standard Deviation of 0.20 1) Develop own protocol for Table 2.5: BCA Assay i. Label 62 microcentrifuge tubes ii. Make 0.5ug/ul BSA standard (250ul of 2mg/ml BSA + 750ul H20)
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iii. Do the same thing as above with 0.5ug/ul lysozyme and 0.5ug/ul gelatin iv. Make about 200ul each of undiluted, ½ diln, ¼ diln, 1/10 diln, 1/20 diln, 1/40 diln of the unknown protein v. Add water into tubes #1-54, then add samples, close and mix
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  • Spring '08
  • Fairclough
  • Assay, Bradford protein assay, Bicinchoninic acid assay, Bradford Assay, unknown protein, original unknown protein

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