In Affiab's laboratory research, various types of antibodies are used for different applications, including Western blotting, chromatin immunoprecipitation (ChIP), immunohistochemistry (IHC), and flow cytometry (FACS). Here are some common types of antibodies used in these techniques:
Polyclonal Antibodies:
Application: Western blotting, ChIP, IHC, FACS
Description: Polyclonal antibodies are produced by multiple B cell clones, each producing antibodies against different epitopes of the target protein. They recognize various sites on the target protein, providing robust signals.
Monoclonal Antibodies:
Application: Western blotting, ChIP, IHC, FACS
Description: Monoclonal antibodies are produced by a single B cell clone, recognizing a specific epitope on the target protein. They offer high specificity and reproducibility, making them ideal for many applications.
Primary Antibodies:
Application: Western blotting, ChIP, IHC, FACS
Description: Primary antibodies directly bind to the target protein. They are used in conjunction with detection methods to visualize and quantify the presence of the target protein.
Secondary Antibodies:
Application: Western blotting, ChIP, IHC, FACS
Description: Secondary antibodies do not directly bind to the target protein but instead bind to the primary antibody. They are conjugated to enzymes or fluorophores for signal detection. Secondary antibodies are species-specific to the host animal of the primary antibody.
Conjugated Antibodies:
Application: IHC, FACS
Description: Antibodies can be conjugated to enzymes (for colorimetric detection in IHC) or fluorophores (for fluorescence detection in IHC and FACS). This allows for visualization and quantification of the target protein.
Isotype Controls:
Application: FACS
Description: Isotype controls are antibodies with the same isotype as the primary antibody but specific for an irrelevant antigen. They are used as controls to distinguish specific staining from non-specific background in flow cytometry experiments.
Tag-specific Antibodies:
Application: Western blotting, ChIP
Description: Antibodies specific to tags (e.g., FLAG, Myc, HA) added to recombinant proteins for easier detection and purification.
AffiAb chose these antibodies carefully based on the specific requirements of the experiment and the characteristics of the target protein. Additionally, optimization and validation of antibodies are crucial for obtaining reliable and reproducible results.