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Looking for Monoclonal Antibodies?
Monoclonal Antibodies for Western Blot Analysis :
- A Monoclonal antibody provides higher specificity, purity and consistency than polyclonal antibodies, resulting in lower backgrounds in western blot results.
Monoclonal Antibody Services :
- Looking to create specific antibodies for your protein of interest? There are many custom services which can create a monoclonal antibody for you in several weeks to months.
There are several ways of making monoclonal antibodies:
- If you have purified protein, simply give this protein for custom monoclonal antibody services.
- If you do not have purified protein, use Peptide Antibody Production Services.
How Much Antigen or Protein do I need for Monoclonal Antibody Development?
For Peptides as Antigens:
- 10 mg is generally enough
- Less peptide may be needed if your peptide is highly purified
- If you need affinity purification of an antibody, around 15 mg of peptide may be needed in dry form.
For Protein Antigens:
- 2-5 mg’s of protein antigen at a concentration of 0.5-1 mg/mL are usually needed.
- Indicate the buffer used and the protein concentration.
- Protein should be 3 – 5 mg at a concentration of 0.5 mg/ml or more.
- For antibody affinity purification, 5 mg of soluble protein is needed to prepare an immunoaffinity column.
Characteristics of the Monoclonal Antibody:
- Specific – due to their nature of originating from one clone specificity is the same for every antibody
- Isotype – all antibodies (and monoclonal antibody batches) are identical giving reproducibility
- Affinity for antigen – monoclonal antibodies
Applications of Monoclonal Antibodies :
- Western Blot – Monoclonal provides better results and less background usually
- Cell Sorting (cell isolation), FACS, or Cell depletion
- Confocal Microscopy and Imaging
- Protein purification
- Diagnosing diseases such as cancer or infectious diseases – monoclonals are also used as therapeutic molecules
Benefits of the Monoclonal Antibody:
Monoclonal antibodies are excellent as the primary antibody in an assay due to their great specificity. Monoclonals recognize only one epitope of the antigen and are highly specific to that particular antigen, and thus will usually give substantially less background staining than polyclonal antibodies. . When compared to the heterogeneity of polyclonal antibodies, the homogeneity of monoclonal antibodies is very high. If experimental conditions are kept constant, using monoclonal antibodies will make results consitently reproducible between experimental replicates due to the clonal nature of monoclonal antibodies. Their specificity also makes them extremely efficient for the binding of antigen within a mixture of related molecules, such as in the case of affinity purification.
Monoclonal antibodies (mAb) are a single type of antibody that are identical and are directed against a specific epitope (antigen, antigenic determinant) and are produced by B-cell clones of a single parent or a single hybridoma cell line. A hybridoma cell line is formed by the fusion of a one B-cell lymphocyte with a myeloma cell. Some myeloma cells synthesize single mAb antibodies naturally.