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Antigens
Antigens for antibody production
Here you get some information about antigens used for antibody production. Which kind of antigens can be used and what amount and conditions are needed for the immunizations.

Antigens


General Information

Definition

Antigens (antibody generating) are molecules where lymphocytes and antibodies can bind. It is the molecule you want to generate the antibodies against. The part of the antigen, where the antibody binds is called epitope. Lymphocytes can detect nearly any kind of antigens. This ensures the somatic recombination of the lymphocytes. The antibody-encoding genes are recombinated to generate a vast amount of different antibodies, recognizing any kind of molecules.

Difference between native proteins and peptides as antigen

Generating antibodies from peptides will lead to antibodies that recognize the peptide. It is likely that the final antibodies will recognize the native protein, but there is no guarantee. The three-dimensional structure or modifications like glycosylation and surface lipids can block the epitope in the native protein. You can raise your chance to get good peptide-antibodies by using more than one peptide. Antibodies generated from the native protein leads to antibodies recognizing multiple epitopes of the protein. This leads to a higher chance that the final antibody pool recognize the native protein, but also increase the chance that a cross-reactivity with other proteins occur.
Peptide Section

Easy to produce Proteins

Background

You need an antibody against a recombinant protein or a protein, that you have in your stock. Just send us the protein on ice, dry ice or lyophilized and we start the production of the antibodies as soon as the antigen arrives.

Requirements

  • Protein concentration: 0.3 - 2 mg/ml
  • Total amount for the immunization of one animal: > 0.2 mg for proteins and > 1 mg for peptides
  • Total amount for the affinity purification: At least 0.5 mg total protein
  • Buffer: 1 x PBS, 0.9% NaCl or other physiological non-toxic buffers
We can handle antigens that don't hit the requirements. We are able to introduce toxic antigens in liposomes, to increase the concentration or to immunize with a smaller amount of antigen. Please contact us for more information.

Proteins from a SDS-Gel

Background

You need an antibody against a protein that is hard to purify or from a protein that is included in a SDS-Gel.

SDS-Gel Electroelution

Electroelution from SDS-PAGE proteins When you are not able to purify the protein in large amounts or in a native way, you can put 100 µg protein on three lanes of a standard SDS-Gel and let it run. Dye it with coomassie and cut your protein band out of the gel. Send it without the addition of any water/solutions at room temperature. We elute the protein from the gel and use it for immunizations.


Requirements

  • Load as much protein into one SDS-Gel lane as possible
  • Load at least 0.3 mg protein in the lanes of one SDS-Gel
  • Run the gel and dye it with coomassie
  • Cut your protein band from the gel
  • Send us the gel pieces in a 2-ml or 15-ml tube
  • Do not add water or any solvents. Just send us the gel as it is.

Peptide Antibodies

Background

You have a protein that you can't express, produce or buy. You can't purify the protein in a native way. You only know the sequence from the protein.

Peptide antibodies

Send us the protein sequence. We suggest you synthetic peptides that represent the protein. The peptides are conjugated to enhance the antigenicity. We synthesize the peptides, perform the conjugation and use them for the immunizations. For more information on peptide antibodies:
Peptide Antibodies

Requirements

  • Peptide concentration: 0.3 - 2 mg/ml
  • Total amount for the immunization of one animal: > 1 mg for peptides
  • Total amount for the affinity purification: Approximately 3 mg total
  • Buffer: 1 x PBS, 0.9% NaCl or other physiological non-toxic buffers
We can handle antigens that don't hit the requirements. We are able to introduce toxic antigens in liposomes, to increase the concentration or to immunize with a smaller amount of antigen. Please contact us for more information.

DNA, Small Molecules, other antigens

Background

You want to produce antibodies against a virus particle, DNA, small molecules, drugs or other substances.

DNA as antigen

We successfully performed DNA immunization. Please ask us for this service.

Small Molecules, Drugs & Virus particles

Information that helps us to produce antibodies against small molecules are the following:
  • Endotoxin concentration
  • Toxicity of the molecule
  • LD50
  • Did you try to produce antibodies against this molecule, were there some difficulties?
  • Are the molecules inactivated/active?

Whole cells

We need the following information of your whole cells:
  • Are the cells pathogenous?
  • Are the cells sterile?
  • Cell concentration


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