Altman:Protocols/Preparing Reagents/Making GOC: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
David Altman (talk | contribs) No edit summary |
David Altman (talk | contribs) No edit summary |
||
| Line 1: | Line 1: | ||
{Template:Altman}} | {{Template:Altman}} | ||
<div style="padding: 10px; width: 700px; border: 5px solid #B22222;"> | <div style="padding: 10px; width: 700px; border: 5px solid #B22222;"> | ||
| Line 39: | Line 39: | ||
2. Make 43 mg/mL glucose oxidase (GO) in 2xAB. | 2. Make 43 mg/mL glucose oxidase (GO) in 2xAB. | ||
* Combine 17 mg of GO with 400 μL 2xAB to make ~400 μL of GO. | \indent * Combine 17 mg of GO with 400 μL 2xAB to make ~400 μL of GO. | ||
3. Make 7.2 mg/mL catalase. | 3. Make 7.2 mg/mL catalase. | ||
Revision as of 22:47, 14 November 2012
Department of Physics, Willamette University
Making Glucose Oxidase Catalase
FINAL CONCENTRATION of 50x GOC stock: 10.8 mg/mL glucose oxidase, 1.8 mg/mL catalase in 1x AB-DTT and 50% Glycerol
- Glucose Oxidase, Aspergillus niger (Calbiochem, 346385)
- Catalase, from Bovine Liver (Sigma, 19.9 mg/mL)
- Glycerol, Anhydrous
1. Make 2x Assay Buffer (2xAB)
| 1 mL 2xAB | |
|---|---|
| 10x AB stock | 200 uL |
| 100x DTT stock | 20 uL |
| ddwater | 780 uL |
2. Make 43 mg/mL glucose oxidase (GO) in 2xAB. \indent * Combine 17 mg of GO with 400 μL 2xAB to make ~400 μL of GO.
3. Make 7.2 mg/mL catalase.
- Combine 145 μL of catalase with 255 μL of 2xAB to make 400 μL of catalase.
4. Add glycerol to the GO and catalase stocks 1:1 volume/volume.
- Add 400 μL of glycerol to each, being careful to not introduce bubbles, especially in the GO.
- This produces 100x stocks of GO and catalase.
5. Combine the GO stock and catalase stock 1:1 volume/volume to yield a 50x stock of GOC.
6. Aliquot into 50 μL stocks, which are stored at 20°C
