9. 1Â SDS-PAGE running buffer: 25 mM Tris–HCl pH 8.3,
192 mM glycine, 0.1% SDS. Measure 100 mL of 10Â
SDS-PAGE buffer into a 1 L measuring cylinder, add 900 mL
of deionized water, and then mix thoroughly.
10. 10Â transfer buffer: 250 mM Tris–HCl, 2 M glycine. Weigh
30.0 g of Tris-base and 144.1 g of glycine into a glass beaker
and dissolve with 500 mL of deionized water using a magnetic
stirrer. After all the chemicals have dissolved completely, adjust
the volume to 1 L with deionized water and mix thoroughly.
The solution can be kept at room temperature.
11. Methanol (analytical grade).
12. 1Â transfer buffer: 25 mM Tris–HCl, 200 mM glycine, 20%
methanol. Measure 700 mL of deionized water into a 1 L
measuring cylinder, then add 100 mL of 10Â transfer buffer
followed by 200 mL of methanol and then mix thoroughly.
This solution must be freshly prepared and cooled to 4
C
before use.
13. 10Â TBS buffer: 200 mM Tris–HCl, 1.4 M NaCl. Weigh
24.2 g of Tris base and 80 g of NaCl into a glass beaker and
dissolve with 800 mL of deionized water using a magnetic
stirrer. After the chemicals have dissolved completely, adjust
the pH to 7.6 with concentrated HCl (see Note 6) followed by
adjusting the volume to 1 L with deionized water and mixing
thoroughly. The solution can be kept at room temperature.
14. 0.1% Ponceau S (w/v) in 5% (v/v) acetic acid.
15. Plastic box with a lid.
16. 1Â TBS-T: 20 mM Tris–HCl, 140 mM NaCl, 0.05% Tween
20. Measure 100 mL of 10Â TBS buffer into a 1 L measuring
cylinder, then add 900 mL of deionized water and 500 μL of
Tween 20 and mix thoroughly.
17. 5% skim milk: 20 mM Tris–HCl, 140 mM NaCl, 0.05% Tween
20, 5% dry skim milk. Weigh 5 g of skim milk powder into a
glass beaker and dissolve with 100 mL of 1Â TBS-T buffer
using a magnetic stirrer. The solution can be kept for 1 week at
4
C.
18. Precision Plus Protein™ All Blue Prestained Protein
Standards.
19. Immobilon
® Forte Western HRP Substrate.
20. Azure C400 imaging system.
21. 100% isopropanol.
22. Mouse monoclonal antibody HB112 (see Note 10).
23. Goat anti-mouse IgG horseradish peroxidase (HRP) conjugated secondary antibody (see Note 11).
24. 3MM Chr cellulose chromatography paper.
Zika Virus-Like Particles
189
192 mM glycine, 0.1% SDS. Measure 100 mL of 10Â
SDS-PAGE buffer into a 1 L measuring cylinder, add 900 mL
of deionized water, and then mix thoroughly.
10. 10Â transfer buffer: 250 mM Tris–HCl, 2 M glycine. Weigh
30.0 g of Tris-base and 144.1 g of glycine into a glass beaker
and dissolve with 500 mL of deionized water using a magnetic
stirrer. After all the chemicals have dissolved completely, adjust
the volume to 1 L with deionized water and mix thoroughly.
The solution can be kept at room temperature.
11. Methanol (analytical grade).
12. 1Â transfer buffer: 25 mM Tris–HCl, 200 mM glycine, 20%
methanol. Measure 700 mL of deionized water into a 1 L
measuring cylinder, then add 100 mL of 10Â transfer buffer
followed by 200 mL of methanol and then mix thoroughly.
This solution must be freshly prepared and cooled to 4
C
before use.
13. 10Â TBS buffer: 200 mM Tris–HCl, 1.4 M NaCl. Weigh
24.2 g of Tris base and 80 g of NaCl into a glass beaker and
dissolve with 800 mL of deionized water using a magnetic
stirrer. After the chemicals have dissolved completely, adjust
the pH to 7.6 with concentrated HCl (see Note 6) followed by
adjusting the volume to 1 L with deionized water and mixing
thoroughly. The solution can be kept at room temperature.
14. 0.1% Ponceau S (w/v) in 5% (v/v) acetic acid.
15. Plastic box with a lid.
16. 1Â TBS-T: 20 mM Tris–HCl, 140 mM NaCl, 0.05% Tween
20. Measure 100 mL of 10Â TBS buffer into a 1 L measuring
cylinder, then add 900 mL of deionized water and 500 μL of
Tween 20 and mix thoroughly.
17. 5% skim milk: 20 mM Tris–HCl, 140 mM NaCl, 0.05% Tween
20, 5% dry skim milk. Weigh 5 g of skim milk powder into a
glass beaker and dissolve with 100 mL of 1Â TBS-T buffer
using a magnetic stirrer. The solution can be kept for 1 week at
4
C.
18. Precision Plus Protein™ All Blue Prestained Protein
Standards.
19. Immobilon
® Forte Western HRP Substrate.
20. Azure C400 imaging system.
21. 100% isopropanol.
22. Mouse monoclonal antibody HB112 (see Note 10).
23. Goat anti-mouse IgG horseradish peroxidase (HRP) conjugated secondary antibody (see Note 11).
24. 3MM Chr cellulose chromatography paper.
Zika Virus-Like Particles
189