WB & IP Positive Hybridoma Screening Introduction

Western blotting (WB) and immunoprecipitation (IP) assays rely heavily on high-quality antibodies, which determine the success or failure of experiments. Therefore, targeted hybridoma screening is critical in the production of antibodies from hybridomas, and Creative Biolabs offers a comprehensive hybridoma platform that can provide you with precise hybridoma screening services.

How to Choose Antibodies for WB?

WB is also known as the protein blotting method. The basic principle of WB is to color a gel electrophoresis-treated cell or biological tissue sample with a specific antibody. Information on the expression of a specific protein in the cell or tissue under analysis is obtained by analyzing the location and depth of the coloring.

In WB, the selection of antibodies is critical to the success or failure of the experiment. When choosing a primary antibody, we need to make the most scientific choice considering the specific experimental conditions. The factors to be considered generally include the information on the target protein, the species of the sample, and the choice of monoclonal or polyclonal antibody.

The target protein: Determine the appropriate antibody according to the target protein. It is important to note that some proteins have different isoforms, so it is necessary to determine the specific isoform before selecting an antibody to accurately select the appropriate antibody.
Species information of the experimental sample: When selecting antibodies, it is necessary to select antibodies based on the species information of the sample to achieve the desired effect. The antibodies are generally internally validated against common target samples of different genera before leaving the factory to determine the different genus types that can be satisfied. Also, the relevant information will be displayed in the species-reactivity of the product specification.
Monoclonal & polyclonal antibodies: In antigen-antibody reactions, monoclonal antibodies are generally specific but have a relatively small affinity and a relatively low sensitivity for antigen detection. In contrast, polyclonal antibodies are slightly less specific, but the affinity of the antibody is strong and the sensitivity is high.

What Is Immunoprecipitation?

IP is a method that uses an antigen-antibody-specific reaction to purify and enrich a target protein. The basic principle of IP is to add specific antibodies and Protein A/G-Beads (Protein A/G immobilized on magnetic beads) to a cell lysate containing the target antigen. The "Antigen-Antibody-Protein A/G-Beads" complex is formed based on the specificity of the antigen and antibody and the FC of Protein A/G and antibody. Unbound heteroproteins are removed from the solution by washing and centrifugation, and the solution is finally tested for the presence of the target protein.

When selecting a primary antibody for IP experiments, it is also necessary to determine the appropriate antibody based on the target protein, sample species information, and other conditions. If no corresponding antibody is available, IP experiments can be performed by using an epitope to label the protein and then selecting a labeled antibody against this epitope.

WB & IP-Positive Hybridoma Screening

When screening WB&IP-positive hybridomas, antibody performance, and antibody specificity testing are required. With a well-defined testing process, the performance and specificity of the antibody can be confirmed to ensure the accuracy and success of subsequent experiments. In general, IHC & IF-positive hybridomas can often be used for WB or IF as well, so adding IHC-positive hybridomas or IF-positive hybridomas at screening will greatly increase the chances of success.

Creative Biolabs is willing to share the key points and experience of WB&IP-positive hybridoma screening with you to promote the development of your project.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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