CASLO Laboratory ApS, Technical University of Denmark, Diplomvej 381, DK-2800 Lyngby, Denmark
Tel: (+45) 70 23 28 60  Fax: (+45) 70 23 29 90  E-mail:info@caslo.com

Polyclonal Antibody Services
antibody-2.jpg

CASLO Laboratory offers a range of polyclonal antibody services according to customers requirements. Rabbits and other animals can be immunized with peptides or recombinant proteins and  CASLO Laboratory offers complete immunization packages at very competitive prices. Antibodies are delivered with detailed product report and titer guaranty. Please send an e-mail with your request and we will get back to you with a detailed project description and price quotation within 24 hours.

  • Only animals breeded for immunization are used, and animals are kept at conditions which minimize risk for interference with pathogens.
  • CASLO Laboratory can assist with identification of the correct amino acid sequence for immunization. 
  • CASLO Laboratory offers a complete peptide immunization package with peptide synthesis, peptide purification to a purity higher than 85%, conjugation to KLH and BSA carrier proteins, immunization of two New Zeeland White Rabbits (or other animals), isolation of preimmune and specific sera, ELISA screening against peptide-BSA conjugate, delivery of lyophilized preimmune serum, lyophilized specific serum, free peptide and peptide-BSA conjugate.
  • CASLO Laboratory offers a complete recombinant protein immunization package with recombinant protein development, production, isolation and characterization. Immunization of two New Zeeland White Rabbits, ELISA screening against the recombinant protein, delivery of lyophilized preimmune serum, lyophilized specific serum and recombinant protein.
  • Antibodies are delivered with detailed product report and titer guaranty.
  • Possibility for affinity purification or Protein-A purification of sera.
  • Possibility for conjugation of antibodies to enzymes or fluorochromes.
  • Possibility for F(ab')2 fragmentation of antibody.   

 

 

 

Below example is a detailed description of one of CASLO Laboratory's peptide immunization packages: 

  

  • A 17 amino acids long peptide is synthesized representing the protein of interest. Eventually CASLO Laboratory can assist with identifying a linear antigenic sequence to be copied from the protein sequence as a synthetic peptide.
  • The peptide is purified to higher than 85% purity at a total quantity of 25 - 30 mg. Part of the peptide is conjugated to KLH, which is used for immunization. Part of the peptide is conjugated to BSA which is used for ELISA screening (2 mg peptide-KLH conjugate, 5 mg peptide-BSA conjugate and 5 mg free peptide are  delivered together with the serum).
  • Two New Zeeland White Rabbits are prepared for immunization per peptide. Only rabbits breeded specifically for antibody production are used for immunization. The rabbits are kept under conditions which minimize risks for infections.  
  • The following peptide synthesis, immunization and serum preparation protocol is used:
    • Day-1: Synthesis of a 17 amino acids long peptide representing a linear sequence from the protein of interest. At the C-terminus or N-terminus of the peptide will be added cysteine amino acid which will be used for conjugation to KLH and BSA carrier proteins.
    • Day-20: The raw peptide will be purified by HPLC to a purity higher than 85%. The purity will be documented in a HPLC report. The correct sequence and identity of the peptide will be verified by mass spectrometry, and a mass spectrometry report will be enclosed with the peptide.
    • Day 30: Part of the peptide will be conjugated to KLH carrier protein and used for immunization. Part of the peptide will be conjugated to BSA protein and used for ELISA screening (BSA and KLH conjugated peptide will be delivered together with free peptide and serum).   
    • Day-38: Preimmune bleeding of 2-3 mL blood per rabbit (from vein or ear edge). 0.1% sodium azide will be added to the preimmune serum, a small part of the preimmune serum will be used as a negative control in ELISA to verify low unspecific background reaction. The remaining quantity will be aliquoted and stored frozen until lyophilization.
    • Day-40: 1st immunization with 2 mg peptide-KLH conjugate in equal volume complete freunds adjuvans (CFA) per rabbit. The rabbits will be immunized by multipoint subcutaneous injections at the back of the animal.
    • Day-62: 2nd immunization with 1 mg peptide-KLH conjugate in equal volume incomplete freunds adjuvans (IFA). The rabbits will be immunized by multipoint subcutaneous injections at the back of the animal. 
    • Day-76: 3rd immunization with 1 mg peptide-KLH conjugate in equal volume incomplete freunds adjuvans (IFA). The rabbits will be immunized by multipoint subcutaneous injections at the back of the animal.
    • Day-86: 1 - 2 mL test bleeding from vein at the ear edge. Titer will be measured by ELISA and the results will be included in the final report.
    • Day-96: 4th immunization with 1 mg peptide-KLH conjugate in equal volume incomplete freunds adjuvans (IFA). The rabbits will be immunized by multipoint subcutaneous injections at the back of the animal.
    • Day-106: Final bleeding. 0.1% sodium azide will be added to the serum.  
    • Day-107: The blood will be centrifuged refrigerated at 3000 rpm for 15 min and serum will be isolated. A small portion of the serum will be used in ELISA, the remaining serum will be aliquoted in 1 mL portions in polyethylene vials and stored frozen until lyophilization.
    • Day-108: All vials will be lyophilized in freeze dryer.
    • Day-110: One vial will be redissolved in pure water and antibody titer will be measured by ELISA.
    • Day 115: Certificate of analysis will be made with all relevant parameters and ELISA results. Separate certificate of analysis will be prepared for the peptide with all physical parameters, HPLC and mass spectrometry results.
    • Day 118 (approximately four months after receipt of order): The following will be shipped by TNT Express: Approximately 50 - 60 mL serum from two rabbits aliquoted and lyophilized in 1 mL portions. 1 mL lyophilized preimmune serum. 5 mg free lyophilized peptide. 5 mg lyophilized peptide-BSA conjugate. 2 mg lyophilized peptide-KLH conjugate. 2 mg free lyophilized KLH carrier protein. 2 mg free lyophilized BSA carrier protein. Detailed certificate of analysis for peptide and antibody.              

As an additional service CASLO Laboratory offers to purify the rabbit anti-peptide serum by either Protein A or affinity chromatography purification.

 

Please contact CASLO Laboratory for a price quotation or if you need more information.