Antigen Preparation and Synthesis

Almost any antigen may be used to generate antibodies. Polyclonal or monoclonal antibodies are typically produced using proteins, peptides, SDS-Gel fragments, DNA, cells, virus particles, or small molecules such as toxins, haptens, or drugs.

Preparation of Antigens

Some antigens require pretreatment before vaccination, particularly when their antigenicity is low. Many strategies, such as conjugation, liposome introduction, or the use of Davids Cell Preparation KIT, are recommended for improving antigens for vaccinations. The recommended preparation of your antigens is shown in Table #1. The suggested buffers for the antigens are shown in Table #2. If you have any queries about your antigens, please contact us at info@davids-bio.com.

Antigen Selection - What Antigen is the best for my Application

Production of antibodies with your antigen. We start with the immunizations as soon as your customer provided antigen arrives or as soon as the production of your antigen finishes.

1. The protein is available
   When you have purified protein available, the native protein can be used as antigen. The SDS-Gel Extraction service can be used for proteins that are difficult to purify or when the purified protein displays many bands in an SDS-gel. We extract your protein from the SDS-gel pieces for immunization once you cut your specified protein band from the gel.
2. Protein not available
   You can utilize our peptide antibody service when you just know the sequence of your protein. We're aiming for peptides that represent your protein. The antigenicity, solubility, and epitopes of the peptides are identified by our scientists with the help of our own prediction algorithm. This service covers pre-order peptide prediction, synthesis and conjugation.
3. More Antigens for Antibody Generation
   We also provide vaccination using antigens such as DNA, Cells, Virus Particles, Toxins, or Small Molecules. Please take a closer look at how these antigens are handled, as each antigen requires its own preparation.

Antigen

Please take a look at the table to discover which antigens are appropriate for rabbit, chicken, mouse, rat, and guinea pig vaccinations. Furthermore, certain antigens require preparation to improve antigenicity or immune system presentation.

	Antigen	Instructions	Information
	Protein	We use your protein for the immunization	We start the immunizations immediately after your protein arrives
	Protein Expression	We produce your antigen	Expression of your plasmid for the antibody production
	Protein (SDS-Gel)	Cut the gel bands from your protein out of the gel	Save expensive and time consuming purification steps
	Peptides	We can synthesize and conjugate your peptides or use your ones	Let us produce your peptide or send us your peptide for the conjugation and immunization
	Cells	Use our cell immunization kit to prepare for immunizations	Send us your cells prepared with our cell immunization KIT
	DNA	We prepare your DNA for the immunizations	Send us your DNA and we prepare it for direct immunization
	Small Molecules	We prepare your small molecules for the immunization	Please contact us when you want to send small molecules

Buffer Requirements

You may use any non-toxic buffer you prefer. Please consult the table of substance concentrations to determine which buffers are appropriate for vaccinations. The maximum concentrations in the table are for antigens at 1 mg/ml concentration.

Buffer Conditions	Maximum Concentration
Urea	2 M
NaCl	200 mM
Imidazole	200 mM
Dithiothreitol (DTT)	2 mM
Citrate or Acetate	100 mM
Glycerol	10%
SDS	0.1%
EDTA	1 mM
TRIS	100 mM

Buffer
Triton X-100

Additional Information

As part of our quality control for our antibody services, we perform ELISA titer determinations. The ELISA values may differ from the true titer when using proteins isolated from SDS-gel or cells as antigen. Without adapting for the individual cell line, not every cell binds to ELISA plates. The same applies equally for SDS-coated denaturated proteins. If you have any more questions, please contact us.