Thermo Scientific Pierce Protein Assay Technical Handbook Version 2
Thermo Scientific Pierce Protein Assay Technical Handbook Version 2
Thermo Scientific Pierce Protein Assay Technical Handbook Version 2
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<strong>Thermo</strong> <strong>Scientific</strong> Total <strong>Protein</strong> <strong>Assay</strong>s<br />
Preparation of Standards<br />
Use this information as a guide to prepare a set of protein standards.<br />
Dilute the contents of one Albumin Standard (BSA) ampule<br />
into several clean vials, preferably using the same diluent as the<br />
sample(s). Each 1mL ampule of 2.0mg/mL Albumin Standard is<br />
Preparation of Diluted Albumin (BSA) Standards for BCA <strong>Assay</strong>, BCA Reducing<br />
Agent-Compatible <strong>Assay</strong> and <strong>Pierce</strong> 660nm <strong>Assay</strong>.<br />
Dilution Scheme for Standard Test Tube Protocol and Microplate Procedure<br />
(Working Range = 20-2,000µg/mL)<br />
Vial<br />
Volume<br />
of Diluent<br />
Volume and<br />
Source of BSA<br />
Final BSA<br />
Concentration<br />
A 0 300µL of stock 2,000µg/mL<br />
B 125µL 375µL of stock 1,500µg/mL<br />
C 325µL 325µL of stock 1,000µg/mL<br />
D 175µL 175µL of vial B dilution 750µg/mL<br />
E 325µL 325µL of vial C dilution 500µg/mL<br />
F 325µL 325µL of vial E dilution 250µg/mL<br />
G 325µL 325µL of vial F dilution 125µg/mL<br />
H 400µL 100µL of vial G dilution 25µg/mL<br />
I 400µL 0 0µg/mL = Blank<br />
sufficient to prepare a set of diluted standards for either working<br />
range suggested. There will be sufficient volume for three<br />
replications of each diluted standard.<br />
Preparation of <strong>Protein</strong> Standards for Coomassie Plus (Bradford) <strong>Assay</strong> and<br />
Coomassie (Bradford) <strong>Assay</strong>.<br />
Dilution Scheme for Standard Test Tube and Microplate Protocols<br />
(Working Range = 100-1,500µg/mL)<br />
Vial<br />
Volume<br />
of Diluent<br />
Volume and<br />
Source of BSA<br />
Final BSA<br />
Concentration<br />
A 0 300µL of stock 2,000µg/mL<br />
B 125µL 375µL of stock 1,500µg/mL<br />
C 325µL 325µL of stock 1,000µg/mL<br />
D 175µL 175µL of vial B dilution 750µg/mL<br />
E 325µL 325µL of vial C dilution 500µg/mL<br />
F 325µL 325µL of vial E dilution 250µg/mL<br />
G 325µL 325µL of vial F dilution 125µg/mL<br />
H 400µL 100µL of vial G dilution 25µg/mL<br />
I 400µL 0 0µg/mL = Blank<br />
Dilution Scheme for Enhanced Test Tube Protocol<br />
(Working Range = 5-250µg/mL)<br />
Vial<br />
Volume<br />
of Diluent<br />
Volume and<br />
Source of BSA<br />
Final BSA<br />
Concentration<br />
A 700µL 100µL of stock 250µg/mL<br />
B 400µL 400µL of vial A dilution 125µg/mL<br />
C 450µL 300µL of vial B dilution 50µg/mL<br />
D 400µL 400µL of vial C dilution 25µg/mL<br />
E 400µL 100µL of vial D dilution 5µg/mL<br />
F 400µL 0 0µg/mL = Blank<br />
Preparation of Diluted Albumin (BSA) Standards for Micro BCA <strong>Assay</strong>.<br />
Vial<br />
Volume<br />
of Diluent<br />
Volume and<br />
Source of BSA<br />
Final BSA<br />
Concentration<br />
A .45mL 0.5µL of stock 200µg/mL<br />
B 8.0mL 2.0µL of vial A dilution 40µg/mL<br />
C 4.0mL 4.0µL of vial B dilution 20µg/mL<br />
D 4.0mL 4.0µL of vial C dilution 10µg/mL<br />
E 4.0mL 4.0µL of vial D dilution 5µg/mL<br />
F 4.0mL 4.0µL of vial E dilution 2.5µg/mL<br />
G 4.8mL 3.2µL of vial F dilution 1µg/mL<br />
H 4.0mL 4.0µL of vial G dilution 0.5µg/mL<br />
I 8.0mL 0 0µg/mL = Blank<br />
Dilution Scheme for Micro Test Tube or Microplate Protocols<br />
(Working Range = 1-25µg/mL)<br />
Vial<br />
Volume<br />
of Diluent<br />
Volume and<br />
Source of BSA<br />
Final BSA<br />
Concentration<br />
A 2,370µL 30µL of stock 25µg/mL<br />
B 4,950µL 50µL of stock 20µg/mL<br />
C 3,970µL 30µL of stock 15µg/mL<br />
D 2,500µL 2,500µL of vial B dilution 10µg/mL<br />
E 2,000µL 2,000µL of vial D dilution 5µg/mL<br />
F 1,500µL 1,500µL of vial E dilution 2.5µg/mL<br />
G 5,000µL 0 0µg/mL = Blank<br />
Preparation of Diluted Albumin (BSA) for Modified Lowry <strong>Assay</strong>.<br />
Dilution Scheme for Test Tube and Microplate Procedure<br />
(Working Range = 1-1,500µg/mL)<br />
Vial<br />
Volume<br />
of Diluent<br />
Volume and<br />
Source of BSA<br />
Final BSA<br />
Concentration<br />
A 250µL 750µL of stock 200µg/mL<br />
B 625 µ 625µL of stock 40µg/mL<br />
C 310 µ 310µL of vial A dilution 20µg/mL<br />
D 625µL 625µL of vial B dilution 10µg/mL<br />
E 625µL 625µL of vial D dilution 5µg/mL<br />
F 625µL 625µL of vial E dilution 2.5µg/mL<br />
G 800µL 200µL of vial F dilution 1µg/mL<br />
H 800µL 200µL of vial G dilution 0.5µg/mL<br />
I 800µL 200µL of vial H dilution 0µg/mL = Blank<br />
J 1,000µL 0 0µg/mL = Blank<br />
6<br />
For more information, or to download product instructions, visit www.thermoscientific.com/pierce
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