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Custom Polyclonal Antibody Service
Allied's custom polyclonal antibody service department has extensive experience in generating high-titer, high-affinity antisera to a wide variety of antigens. Our staff is ready to assist you in experimental design, laboratory methods, production, purification and conjugation. If you have any questions about experimental designs please contact Dr. James Li (James@alliedbiotechinc.com) via e-mail or by calling him at 301-607-4740.
Below are some Frequently Asked Questions.
- How much antigen is required? Using proprietary immunization methodologies as little as 200 micrograms of antigen can be used for a standard immunization protocol. However, we have been successful using approximately 50 micrograms for valuable antigens. If antigen is abundant immunize with 0.1 to 0.5 mg per injection.
- What is Allied's recommended immunization regime? Customized protocols can always be developed. Otherwise the NIH 90-day standard protocol or 70-day conventional Protocol listed below will be used:
| Subject: | New Zealand White Rabbits |
| Adjuvant: | Complete Freund's Adjuvant (CFA) initially, followed by Incomplete Freund's Adjuvant (IFA) for all subsequent injections. |
| Immunogen: | Typically, proteins and conjugated peptides. Amount of immunogen used for each immunization is 50 - 500 mg. |
| Procedure: | The immunogen is diluted to 1 mL with sterile saline and combined with 1 mL of the appropriate adjuvant. The antigen and adjuvant are mixed thoroughly to form a stable emulsion which is injected subcutaneously and provides enhanced immune reponse from the sustained presence of the immunogen. |
| Bleeds: | Blood is collected from the central ear artery with a 19 gauge needle and is allowed to clot and retract at 37ºC overnight. The clotted blood is then refrigerated for 24 hours before the serum is decanted and clarified by centrifugation at 2500 rpm for 20 minutes. |
| Schedule: | Time 0 Bleed 25 mL (yields 10 mL pre-immune serum). Immunize with antigen in CFA. Week 3: Immunize with antigen in IFA. Week 6: Immunize with antigen in IFA. Week 7: Bleed 50 mL (yields 20 mL serum). Week 10: Immunize with antigen in IFA. Week 11: Bleed 50 mL. |
- How many animals should I put on a study? We recommend two or more animals to allow for immune response variability.
- What type(s) of antigens can be injected? We can immunize animals with native proteins, GST-fusion proteins, peptides, carbohydrates or any other compound of viral, fungal or microbial origin. However, no biohazardous material can be accepted. We recommend the use of Freund's Adjuvant and we can accommodate any requirements that you may have for specialized adjuvants.
- Can you conjugate my peptide to a carrier? Yes. We charge $200 for peptide-carrier protein conjugation to BSA or KLH.
- How should I send the antigen? Antigen can be supplied in a soluble form, as a lyophilizate, a gel slice or on a nitrocellulose membrane. An additional charge of $35 will be applied for gel slice extraction. Please call prior to shipping so that we can prepare to start your project.
- Can you estimate the total cost of my project? Yes. The following guidelines can be used to estimate project costs. Remember, depending on specific instructions you give our staff, actual charges may differ from the estimate. Call for specific estimates or to receive written quotations.
Item |
Subtotal |
Total |
2
rabbits |
$65
each |
$130 |
45-day
duration |
2
x $1.95 per day |
$175 |
2
pre-bleeds |
$20
each |
$40 |
2
test bleeds |
$20
each |
$40 |
100
mL antisera per rabbit |
$1.25
per mL |
$250 |
Here is a list of animals available for polyclonal antibody production and the various charges for the work performed:
| Service | Rabbit | Goat | Sheep | Donkey |
| Cost of animal | $65 | $225 | $225 | Inquire |
| Daily maintenance | $1.95 | $3.45 | $3.45 | $7.00 |
| Pre-Bleed | $20 | $25 | $25 | $25 |
| Injection | No charge | No charge | No charge | No charge |
| Test bleed | $20 | $25 | $25 | $25 |
| Antisera (per mL) | $1.25 | $0.90 | $0.90 | $0.90 |
Please call for pricing of donkey, guinea pig, rat, or mice projects.
© 2003 Allied Biotech, Inc.