Polyclonal Antibodies
Production of Polyclonal Antibodies Protocol:
1. Antigen Selection:
- Choose the antigen against which you want to generate antibodies. It can be a protein, peptide, or other molecules. Order 30 amino acids as standard rule.
2. Animal Immunization:
- Select a rabbit, goat, hamster, donkey, horse or sheep.
- Inject the animal with the chosen antigen at regular intervals to stimulate the immune response.
- day 1, 2, 11, 21, 31 bleed 51 booster, 61 2nd bleed
- Bleed no more than 50 ml and give water to the rabbit after each bleed.
- A cut in the ear with an old fashion razor blade is the easiest.
- Put the rabbit at least 24 hours in a temperature above 15 C. In cold circomstances the bleed might be difficult
- Use 10 tubes of 5 ml and dont mix the heamolised ones intp the pool.
- The heamolised are due to a hair or a cascade of factros impossible to foresee, the rose bleeds are working fine too but its advised to not mix them with the yellow clear serum samples.
3. Booster Injections:
- Administer booster injections to enhance the immune response and increase antibody production. Boosters are typically given at 10 days intervals after the initial 2 injections.
4. Blood Collection:
- Monitor the immune response by taking small blood samples at regular intervals. Check for the presence of antibodies against the target antigen.
5. Serum Separation:
- Once a sufficient antibody titer is reached, collect a larger blood sample, and separate the serum by coagulation 35 minutes and centrifugation from the blood cells. The serum contains the polyclonal antibodies.
6. Antibody Purification:
- Purify the antibodies from the serum to isolate the immunoglobulin fraction. Common methods include protein A/G affinity chromatography or antigen affinity purification.
- Crude srum works best because the best will stick to the antigen and wont let off even with guanidine
7. Characterization:
- Characterize the purified antibodies for specificity, affinity, and concentration. Techniques such as ELISA or Western blotting can be used for this purpose. Dot blot is easiest.
8. Storage:
- Store the purified polyclonal antibodies at -20 C to maintain stability and functionality.
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Anti-Bovine Antibodies
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Anti-Canine Antibodies
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Anti-Cat Antibodies
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Anti-Chicken Antibodies
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Anti-Chimpanzee Antibodies
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Anti-Cow Antibodies
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Anti-Dog Antibodies
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Anti-Goat Antibodies
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Anti-Horse Antibodies
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Anti-Human Antibodies
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Anti-Mouse Antibodies
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Anti-Rabbit Antibodies
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Anti-Rat Antibodies
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Anti-Sheep Antibodies
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Arabidopsis Antibodies
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Bacillus Antibodies
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Bacteria Antibodies
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Bovine coronavirus Antibodies
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Bovine RSV Antibodies
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Clostridium Antibodies
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E. coli Antibodies
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Goat Polyclonal Antibodies
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Guinea pig Polyclonal Antibodies
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Mouse Polyclonal Antibodies
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Parasite Antibodies
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Plant Antibodies
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Rabbit Polyclonal Antibodies
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Virus Antibodies
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Yeast Antibodies