Several techniques are available at Bio-Synthesis to evaluate the quantity, quality,
and function of the antibodies produced. BSI's Protein Analytical Methods can provide you with important and detailed information about polyclonal and monoclonal antibodies and help you throughout the development phase.
Bio-Synthesis can provide you with detailed characterization of your antibody including:
- Antibody titration and quantization by ELISA
- Western Blot Assay
- HPLC, SDS, PAGE IEF Analysis
- Intact mass determination by mass spectrometry of heavy and light chain
- N- and C-terminal sequencing of heavy and light chain by Edman degradation and MALDI MS
To obtain further information, please contact us.
ELISA Development and Testing
ELISAs (Enzyme Linked Immunosorbent Assays) are used to measure the specific binding between an antigen and antibody using a secondary enzyme labeled antibody and substrate system. This is a rapid and sensitive method for evaluating large numbers of monoclonal or polyclonal antibodies at low cost. The scientists at Bio-Synthesis have years of experience developing ELISAs for our clients and our own in-house antibodies; we are pleased to share that experience with you. Once an ELISA has been developed for you, we will transfer the ELISA protocol to you and, if you wish, apply that ELISA to testing of your antibody samples.
Western Blot Assay and Electrophoresis
This technique permits identification of proteins by their molecular weight and their immunoreactivity or the control of the immunoreactivity of serum on the denaturated protein of interest. The sample is prepared and loaded into a pre-cast 10% to 12% Tris-HCl denaturing gel. The standards are included with molecular weights ranging from 6.5kD to 200kD. Two bands are observed after staining in Coomassie Blue rep-resenting the heavy chain (55kD) and the light chain (25kD) of the antibody. Each client is provided with a photo of the gel as an indicator of purity.
Type of sample:
- Purified protein: 15-20 µg
- Cellular lysate containing the protein of interest: 500 µg
Isotype Determination
The isotype of murine antibodies (either in supernatants or ascites) can be determined using murine isotyping kits or strips.
For additional antibody characterization services, visit our proteomic analytical services.