HRP (Spicy Root Peroxidase) marks the antibody process

1.8 mg spicy root peroxidase into a 1 ml glass bottle, add 1 ml of double steamed water dissolved, the liquid brown.

2. Put in a magnetic mixer, electromagnetic stirring and slowly add the newly-made NaIO4 0.2 ml, continue stirring at room temperature 40 min, the liquid appears grassy green.

3. Load all solutions intodialysis bags rinsed repeatedly with deionized water, 4 oC to 1 mM pH 4.4 NaAc dialysis overnight, intermediate fluid change 3-4 times, each 300 ml, the solution is finally light brown.

4. According to the enzyme:antibody s 1:1 ratio prepared antibodies, antibody concentration is not less than 2 mg/ml.

5. to 0.01 M Na2CO3 dialysis overnight, intermediate fluid exchange 3-4 times.

6. Put the dialysis-good HLP solution (light brown) into a glass bottle, add 0.2 M Na2CO3 solution to adjust the pH to 9.0, mix immediately after adding dialysis-good antibodies, while adding electromagnetic"stirring.

< the pH of the > HLP/antibody mixture to 9.0 (between 9-10) with a 0.2 M Na2CO3 solution, and continue stirring at room temperature 3hr.

8. Add the newly made sodium boron hydride solution 100 ul, 4 oC place 2 hr.

< fully dialysis in the "zoom" >9.4 oC, 1×PBS, every 2hr change of fluid, a total of 4 times.