"
Team:Calgary/Notebook/Protocols/CoilBindingNitrocelluloseAssay
From 2013.igem.org
(Difference between revisions)
TRemondini (Talk | contribs) |
Sharonfeng1 (Talk | contribs) |
||
| Line 27: | Line 27: | ||
<li>Cut out an appropriately sized piece of nitrocellulose</li> | <li>Cut out an appropriately sized piece of nitrocellulose</li> | ||
<li>Place matching filter paper under the nitrocellulose</li> | <li>Place matching filter paper under the nitrocellulose</li> | ||
| - | <li>Add 5 | + | <li>Add 5 μL of sample to (Prussian blue ferritin, ferritin, negative BSA control) to each wanted spot</li> |
<li>Let the nitrocellulose dry for 15 minutes</li> | <li>Let the nitrocellulose dry for 15 minutes</li> | ||
| - | <li>Add 7.5 | + | <li>Add 7.5 μL of substrate solution (1:1:1 ratio of 30% hydrogen peroxide, 10 mg/mL ABTS or TMB, and 0.2 M pH 3.6 acetate buffer)</li> |
<li>Record results via photographs every two minutes</li> | <li>Record results via photographs every two minutes</li> | ||
</ol> | </ol> | ||
Latest revision as of 00:35, 28 September 2013
Nitrocellulose Assay
Coil Binding Nitrocellulose Assay
- 10 mg/mL TMB
- 0.2 M pH 3.6 Acetate buffer
- 30% Hydrogen Peroxide
- Nitrocellulose
- Filter Paper
- 0.022 mg/mL Prussian Blue Ferritin
- 1 mg/mL Bovine Serum Albumin
- 0.5 mg/mL Recombinant Prussian Blue Ferritin
- 5% Skim Milk solution
- 1x 0.1% TBS-Tween
Protocol
- Cut out an appropriately sized piece of nitrocellulose
- Place matching filter paper under the nitrocellulose
- Add 5 μL of sample to (Prussian blue ferritin, ferritin, negative BSA control) to each wanted spot
- Let the nitrocellulose dry for 15 minutes
- Add 7.5 μL of substrate solution (1:1:1 ratio of 30% hydrogen peroxide, 10 mg/mL ABTS or TMB, and 0.2 M pH 3.6 acetate buffer)
- Record results via photographs every two minutes
