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FAQ |
How
many animals should I use?
What species should I use?
How should the immunogen be delivered?
When will I obtain my first serum sample:
How much antigen do I need to make a good antibody?
Do I need to conjugate my immunogen?
Do I still need an adjuvant if my peptide is conjugated?
Does my serum need to be purified?
What kind of purification should I perform?
What is your shipping address?
What is your post office address?
What is your FAX number?
Do you accept credit cards?
How many animals should I use?
We suggest at least 2 animals per project. This will allow for some individual variability between animals. If you are looking for high volumes of serum, please contact us.
What species should I use?
In many cases, rabbits are usually the first choice for polyclonal antibodies for a research lab, but other species should be considered. There are a number of advantages and reasons for using sheep or goats to produce antibodies.
How should the immunogen be delivered?
Most times, the immunogen will be delivered in a solution or suspension and should be stored at -20 C or colder. The greatest concern we have while handling immunogen is to keep the degradation process to a minimum. The easiest suggestion we have is to keep the number of freeze/thaw cycles to a minimum. At Capralogics, we suggest that the investigator aliquot the immunogen prior to shipping it. This will save a freeze thaw cycle at Capralogics thus help preserve the integrity of the immunogen. If the immunogen arrives in bulk, we will aliquot it into pre-determined volumes and store it at the required temperature.
When will I obtain my first serum sample?
The date of the first serum sample is determined at the time we initiate a project. Without any specific request, we normally produce the first test bleed at day 52. A common request from researchers is to take a sample at day 35, and we are most willing to accommodate. A good ELISA at day 35 means the antibody looks very promising. A weak ELISA at day 35 does not necessarily mean the final product will be mediocre. Novel antibodies take time to develop, and in many cases the animals will need a longer period of time to produce an adequate titer.
How much antigen do I need to make a good antibody?
Typically between 1-2 mg per rabbit is sufficient if the antigen is immunoreactive. A goat or sheep may require between 2-4 mg. This would be enough to last at least 6 months. For formulation reasons, we prefer the antigen to be at least 1 mg/mL in concentration.
Do I need to conjugate my immunogen?
Conjugation makes the immunogen more immunogenic by making it physically large enough for the immune system to recognize it. We recommend that small peptides (less than 13-15 KD or less than 25-27 amino acids) be conjugated. During conjugation your peptide is attached to a carrier protein making for a larger and more immunogenic molecule that is coated with your peptide. The larger molecule allows the immune system to recognize it easier. We have great success using keyhole limpet hemocyanin (KLH). Other good carrier proteins are bovine serum albumin (BSA) and ova albumin (OVA)
Do I still need an adjuvant if my peptide is conjugated?
Yes. The adjuvant help stimulate the immune system and helps in the delivery of the immunogen.
Does my serum need to be purified?
Not always. The main reason for purification is to remove any non-specific staining or multiple bands on Western Blots. Another reason for purification is to concentrate the antibody to increase signal strength.
What kind of purification should I perform?
Traditionally, an affinity purification (against the antigen) is the purification of choice. The end result is an extremely clean antibody. Like most things, there is a catch though. Sometimes some of the most highly avid antibodies may not detach from the affinity matrix, thus cannot be recovered. One must not rule out purifying against protein G or protein A, or an ammonium sulfate precipitation. Many times, these will prove sufficient.
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| Needless to say, most people want the best antibodies in the shortest amount of time, but one must remember we are working with a natural process and a wide variety of proteins that will react differently. A good response at day 31-35 is usually a good indicator of future success, but one must be patient. Many times the initial test bleed will react well against the peptide, but good reactivity against the native protein usually will take more time to develop. Be patient. This is where antibody production has much in common with wine making. |
Shipping
Address:
Capralogics,
Inc.
235 Czeski
Road
Hardwick,
MA 01037 USA
Post
Office Address:
Capralogics,
Inc.
PO Box
356
Hardwick,
MA 01037 USA
FAX: 413-643-0067
We Accept Credit Cards for Payment
Phones: 413-477-6866 or 800-975-6866
