Polyclonal Antibodies

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.

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.

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.

Monitor the immune response by taking small blood samples at regular intervals. Check for the presence of antibodies against the target antigen.

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.

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

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.

Store the purified polyclonal antibodies at -20 C to maintain stability and functionality.

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