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| To prepare serum from each bleed, simply allow the blot to clot and leave at room temperature overnight. The next morning, centrifuge the cells in an SS34 rotor at 10,000 rpm (12,000 X g) for 5 min. Aliquot the serum and store it at -80°C. |
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1. Pre-bleed approximately 10 ml of blood from the rabbit.
2. Inject 200 μg to 1 mg of the desired antigen in 1 to 2 ml of 1:1 PBS:Freund's Adjuvant Emulsion.
3. Inject the rabbit at various sites subcutaneously along its back, and intramuscularly in its thigh.
4. Three weeks after the initial injections, boost by injecting antigen in incomplete Freund's Adjuvant.
5. Two weeks after the first boost, perform another boost with antigen in incomplete Adjuvant.
6. 10 days later, bleed the rabbit and repeat the boosting injection.
7. 10 days later, bleed the rabbit and repeat the boosting injection.
8. The rabbit can be kept indefinitely and when further bleeds are required, two boosts 10 days apart should be sufficient to generate a lot of antibody.
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| PBS |
| pH 7.2
2.7 mM KCl
4.3 mM Sodium Phosphate Dibasic (Na2HPO4)
137 mM NaCl
1.8 mM Potassium Phosphate Monobasic (KH2PO4)
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Potassium Chloride
Freund's Adjuvant
Sodium Chloride
Potassium Phosphate, Monobasic
Sodium Phosphate, Dibasic
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No hints are associated with this bioProtocol
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