B Cell-based Western Blot Assay

Western blot (WB) is a widely used technique to isolate and identify specific proteins in a protein mixture extracted from cell or tissue lysates based on the merits of specific antibodies. Creative Biolabs offers the highly sensitive WB assay for clients to identify antigenic peptides within a tumor antigen using target protein-specific antibodies.

Introduction of Western Blot

Fig.1 Using western blot to separate and detect allergens.¹

In this technique, protein samples are loaded on electrophoresis gel and separated by molecular weight. The gel is then transferred to PVDF under electricity to produce corresponding bands, as shown on the SDS-PAGE gel. After a blocking step, the membrane is then probed with antigen-specific primary antibody, and bound Abs are detected with enzyme-conjugated secondary antibody. Due to the specificity of the antibody, only one band indicating the targeted antigen can be visible. The targeted protein is quantified by comparing the thickness of the band to a reference protein band.

Western Blot Service at Creative Biolabs

Western blotting is also called immunoblotting and is the most preferred method applied to detect a specific protein antigen in a mixture. Creative Biolabs offers the latest validated materials and reagents for the application of this powerful tool, and we have developed an innovative platform for tumor antigen analysis using this sensitive approach.

Workflow

• Protein Antigen and Epitope Peptide Preparation

Different proteins of the infectious virus or other pathogens can be synthesized for analysis, such as:

• Antibody Preparation

Various sources of polyclonal and monoclonal antibodies are used for epitope analysis.

• Western Blot Analysis

Multiple western blot methods are available to detect and quantify proteins with high-quality antibodies, including:

• Radioactive Western Blot
• Fluorescent Western Blot
• Colorimetric Western Blot

Application of WB in Cancer Epitope Analysis

• Identify the shortest epitope reactive to specific antibodies.
• Provide a comprehensive epitope mapping of a specific antigen.
• Western blot can be used combined with immunoprecipitation to examine the precipitated antigen peptide.

If you have trouble with western blot and want to use this technique in cancer epitope analysis, please contact us and discuss your research with our expert.

Reference

1. Salazar, A.; et al. Allergen-Based Diagnostic: Novel and Old Methodologies with New Approaches. 2017.

For Research Use Only | Not For Clinical Use