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For the Bradford assay, include the concentration of Lysozyme you started with in the header of the table. Series Number S1 [X mg/mL] Lysozyme, mL Buffer, mL Total Vol, mL Dilution Factor [Final ug/mL] Lysozyme 1 10 S2 0.8 10 S3 0.6 10 S4 0.4 10 S5 0.2 10 Sample Prep 1. Prepare dye reagent Coomassie Brilliant Blue G-250 by diluting dye reagent 1:4 with distilled H2O and store in new sterile conical vial with aluminum foil. Store for up to 4 weeks at 4°C. 2. Label your conical tubes appropriately in advance ex) S1.1, S1.2, S1.3 for each triplicate in series 1. You should have 15 tubes in total. - 3. Make a dilution of your 10 mg/mL protein stock. Anything from 0.1 – 1 mg/mL can work, though for our pipetting capacity, 1 mg/mL works best. 4. In your ice bucket you should have a. 0.1 M K Phos pH 7.2 b. Diluted Bradford reagent, wrapped in foil. c. 1 mg/mL protein 5. To a 0.1 ml sample containing 10-100 µg total protein, add 4.9 ml of diluted dye reagent (also remember the blank). These are to be made in triplicate. 6. Invert each ~5 mL sample in a 15 mL conical tube gently to mix. Let samples incubate for ~5 minutes at room temperature (but no longer than 15 minutes) after making and before data acquisition.

For the Bradford assay, include the concentration of Lysozyme you started with in the header of the table. Series Number S1 [X mg/mL] Lysozyme, mL Buffer, mL Total Vol, mL Dilution Factor [Final ug/mL] Lysozyme 1 10 S2 0.8 10 S3 0.6 10 S4 0.4 10 S5 0.2 10 Sample Prep 1. Prepare dye reagent Coomassie Brilliant Blue G-250 by diluting dye reagent 1:4 with distilled H2O and store in new sterile conical vial with aluminum foil. Store for up to 4 weeks at 4°C. 2. Label your conical tubes appropriately in advance ex) S1.1, S1.2, S1.3 for each triplicate in series 1. You should have 15 tubes in total. - 3. Make a dilution of your 10 mg/mL protein stock. Anything from 0.1 – 1 mg/mL can work, though for our pipetting capacity, 1 mg/mL works best. 4. In your ice bucket you should have a. 0.1 M K Phos pH 7.2 b. Diluted Bradford reagent, wrapped in foil. c. 1 mg/mL protein 5. To a 0.1 ml sample containing 10-100 µg total protein, add 4.9 ml of diluted dye reagent (also remember the blank). These are to be made in triplicate. 6. Invert each ~5 mL sample in a 15 mL conical tube gently to mix. Let samples incubate for ~5 minutes at room temperature (but no longer than 15 minutes) after making and before data acquisition.

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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using sample prep aka the protocol use it to fill in the tables

For the Bradford assay, include the concentration of Lysozyme you started with in the header of the table. Series Number S1 [X mg/mL] Lysozyme, mL Buffer, mL Total Vol, mL Dilution Factor [Final ug/mL] Lysozyme 1 10 S2 0.8 10 S3 0.6 10 S4 0.4 10 S5 0.2 10
Transcribed Image Text:For the Bradford assay, include the concentration of Lysozyme you started with in the header of the table. Series Number S1 [X mg/mL] Lysozyme, mL Buffer, mL Total Vol, mL Dilution Factor [Final ug/mL] Lysozyme 1 10 S2 0.8 10 S3 0.6 10 S4 0.4 10 S5 0.2 10
Sample Prep 1. Prepare dye reagent Coomassie Brilliant Blue G-250 by diluting dye reagent 1:4 with distilled H2O and store in new sterile conical vial with aluminum foil. Store for up to 4 weeks at 4°C. 2. Label your conical tubes appropriately in advance ex) S1.1, S1.2, S1.3 for each triplicate in series 1. You should have 15 tubes in total. - 3. Make a dilution of your 10 mg/mL protein stock. Anything from 0.1 – 1 mg/mL can work, though for our pipetting capacity, 1 mg/mL works best. 4. In your ice bucket you should have a. 0.1 M K Phos pH 7.2 b. Diluted Bradford reagent, wrapped in foil. c. 1 mg/mL protein 5. To a 0.1 ml sample containing 10-100 µg total protein, add 4.9 ml of diluted dye reagent (also remember the blank). These are to be made in triplicate. 6. Invert each ~5 mL sample in a 15 mL conical tube gently to mix. Let samples incubate for ~5 minutes at room temperature (but no longer than 15 minutes) after making and before data acquisition.
Transcribed Image Text:Sample Prep 1. Prepare dye reagent Coomassie Brilliant Blue G-250 by diluting dye reagent 1:4 with distilled H2O and store in new sterile conical vial with aluminum foil. Store for up to 4 weeks at 4°C. 2. Label your conical tubes appropriately in advance ex) S1.1, S1.2, S1.3 for each triplicate in series 1. You should have 15 tubes in total. - 3. Make a dilution of your 10 mg/mL protein stock. Anything from 0.1 – 1 mg/mL can work, though for our pipetting capacity, 1 mg/mL works best. 4. In your ice bucket you should have a. 0.1 M K Phos pH 7.2 b. Diluted Bradford reagent, wrapped in foil. c. 1 mg/mL protein 5. To a 0.1 ml sample containing 10-100 µg total protein, add 4.9 ml of diluted dye reagent (also remember the blank). These are to be made in triplicate. 6. Invert each ~5 mL sample in a 15 mL conical tube gently to mix. Let samples incubate for ~5 minutes at room temperature (but no longer than 15 minutes) after making and before data acquisition.
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