Custom Peptide Synthesis, Conjugation, and Anti-Peptide Antibodies

Custom Peptide Synthesis, Conjugation, and Anti-Peptide Antibodies
Anti-Peptide Antibody package is designed to be economical and includes the following:
  • Peptide synthesis (HPLC, Mass Spec, Scale 10-15 mg; 70-85% purity)
  • Supply of free peptide (~ 5 mg is used for antibody production and ELISA and the rest is supplied to the researcher).
  • Conjugation of peptide to carrier protein (KLH, BSA, ovalbumin, etc.).
  • Antibody production in 2 rabbits (or 2 G. Pigs or 2 Chickens, extra) using std. 63 days protocol.
  • ELISA titration of each antibody on first bleed.
  • Approximate duration of projects (from peptide order to first bleed): 9-11 weeks depending upon the peptide length.
  • Free Services: Selection of Antigenic Peptides, BLAST searches, Identification of Signal peptide, Transmembrane domains, Sequence alignments, and other common structural motifs

Peptide Synthesis and Antibody Ordering Information
Peptide Length Std Package Cat # ($$) With Peptide Upgrade Cat # ($$) With Affinity Purification Cat # ($$) With Peptide Amount Upgrade & Aff. Pur. Cat # ($$)
Up to 20 AA ABPEP-20 ABPEP-20U ABPEP-20A ABPEP-20UA
Up to 25 AA ABPEP-25 ABPEP-25U ABPEP-25A ABPEP-25UA
Up to 30 AA ABPEP-30 ABPEP-30U ABPEP-30A ABPEP-30UA
 

Peptide Synthesis and Antibody Ordering Information
Items/ Services Std Package With Peptide Upgrade With Affinity Purification With Peptide Amount Upgrade & Aff. Pur.
Free Protein sequence & antigenicity analyses
Peptides (70-85% pure, mass spec, hplc) 5-10 mg total 10-20 mg total 5-10 mg total 10-20 mg total
Supply of free peptide 5 mg ~10-15 mg ~ 5 mg ~10 mg
Peptide Conjugation (couple ~ 5 mg to KLH etc)
Animals (2 rabbits) & 63-days immunization program
Pre-immune bleeds (2-5 ml/animal)
Production Bleeds (15-20 mls/animal) total 70-80 mls
ELISA Testing of 1st bleeds
Affinity Purification (includes coupling of ~ 5 mg peptide to Sepharose, purification of 1 bleed, ELISA testing of bleeds and supply of the affinity column)    
Antibody Production Guarantee (Minimum antibody titer by ELISA ~1K or repeat the project at no cost (see details below)
 
All projects include 2 rabbits but we can substitute them with 2 G. Pig/Hamsters/or Chicken at no extra charge. Goat and sheep will incur additional charges. We can modify the std immunization and bleed protocols and include more animals if requested. Other animals, not included above, can be provided as well. Please contact ADI for current prices and availability. Std 63 days protocol include up to 5 injections (i.m. and s.c. at multiple sites at 15 days intervals); 1 preimmune bleed (~2 ml per animal), 2 immune bleeds (std. bleeds at weeks 7 and 9; ~15-20 mls per rabbit). All projects may be extended on a monthly basis, for a nominal fee, to collect more antiserum (2 bleeds per month). Please inform us prior to the conclusion of std protocol if you wish to extend the projects. Please consult Some Commonly Asked Questions About Custom Peptide And Antibody Services for bleed volume in other species and optional services.

Peptide Amounts Upgrades: Standard package includes about ~10-20 mg of >90% pure peptide. This is normally sufficient to produce antibodies and have some free peptide (at least 5-10 mg) for additional experiments. However, if affinity purification (need approx. 5-10-mg peptide) is intended or needed, it is a good idea to Upgrade Amounts synthesis (synthesize 20-40 mg of 70-85% pure).

Affinity purification. This package typically includes: Coupling of peptide (up to 10 mg to Sepharose), purification of any 1 bleed of ~ 15 ml or a pool of antiserum up to 15 ml; ELISA testing of both unpurified and affinity pure antibodies, and supply of affinity column. We also monitor the antibody activity in the unbound fraction to make sure antibodies are not overloaded and wasted. Ideally, we need approx. ~10 mg of antigen or peptide for making the affinity matrix. This package (Cat # AFFI-1) can also be ordered separately. Note: Affinity purification is normally scheduled on 2nd bleed (pool of sera from both rabbits) unless antibody titer is poor in the first and 2nd bleed. However, user may elect to delay affinity purification or use just 1 rabbit serum. We must be informed prior to the second bleed.

ELISA. ADI uses proprietary techniques and buffer solutions to coat free peptides on ELISA plates. Therefore antibodies to the carrier protein are not detected. We test individual preimmune and immune serum at 4 dilutions (1:100, 1:1K, and 1:10K, 1:100K). A detailed ELISA report is provided with every test regarding antibody titer of each animal. Antibody titer reported in our ELISA report refers only to anti-peptide antibodies. Complete instructions of performing ELISA and reagents used are given in ELISA Procedure
 
Catalog# Product Description Product Type Antibody Type Size
AFFI-2 Custom Conjugation of peptides/proteins to Agarose (up to 10 mg antigen) Service   1
AFFI-I Custom Affinity Purification of antibodies (Coupling of antigen to Agarose, Ab purification (10 ml), ELISA, and affinity column Service   1