Antibody Applications - Methods, Techniques, Protocols


Antibody Applications




  • Blocking/Neutralization (Blk/Neut) - methods to check antibody specificity or reduce unspecific background staining; used for negative control, competing control, absorption control, isotype control, etc.

  • Chromatin Immunoprecipitation (ChIP) - procedure used to determine whether a given protein binds to or is localized to a specific DNA sequence in vivo.

  • Dot Blot - similar technique as western blotting for detecting proteins in samples that are spotted through circular templates directly onto the membrane.

  • Enzyme Immunoassay (EIA) - An assay that uses an enzyme-bound antibody to detect antigen. The enzyme catalyzes a color reaction when exposed to substrate.

  • Enzyme Linked ImmunoSorbent Assay (ELISA) - technique for detecting the presence of specific substances, such as enzymes, viruses, bacteria, or antibodies in blood.

  • Flow Cytometry (FC) - analysis of cells or subcellular components by detecting fluorescence or light-scatter of sample fractions passing in narrow-stream droplets through a laser beam.

  • Fluorescence Activated Cell Sorting (FACS) - Fluorescence-activated cell-sorting (FACS) is a specialised type of flow cytometry. It provides a method for sorting a heterogenous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell.

  • Functional Assay (FA) - screening or measuring of functionally active proteins present in plasma or culture media using ELISA or other method.

  • Gel Shift (Gel, EMSA) - also called band shift assay, or electrophoretic mobility shift assay (EMSA), a method for detecting DNA-binding proteins.

  • Immunoblotting (IB) - Very small amounts of protein are transferred from gels to nitrocellulose sheets by electrophoresis and then detected by their antibody binding, usually in combination with peroxidase or radioactively labeled IgG. An accurate technique for the specific recognition of very small amounts of protein.

  • Immunocytochemistry (ICC) - technique used to detecting the presence of proteins in cell suspension, cultured cells or cytospin by use of a specific antibody.

  • Immunodiffusion (ID) - technique involving diffusion of antigen or antibody through a semisolid medium, usually agar or agarose gel, resulting in a precipitin reaction.

  • Immuno-Electron Microscopy (IEM) - applying immunohistochemistry or immunocytochemistry method to localize antigens or proteins at sub-cellular level using an electron microscope.

  • Immunofluorescence (IF) - labeling of antibodies or antigens with fluorescent dyes in cells or tissue sections which are visualized using a fluorescence or confocal microscope.

  • Immunohistochemistry (IHC) - localization of antigens or proteins in tissue sections by the use of labeled antibodies as specific reagents through antigen-antibody interactions that are visualized by a marker such as fluorescent dye, enzyme, or colloidal gold.

  • Immunoprecipitation (IP) - procedure by which peptides or proteins that react specifically with an antibody are removed from solution and examined for quantity or physical characteristics.

  • Radioimmunoassay (RIA) - method to measure the amount of a substance using an antibody against the substance for specificity and a radioactive label for detection and measurement.

  • Western Blotting (WB) - technique which transfers proteins from a gel to a nitrocellulose or nylon membrane following electrophoresis and uses specific antibodies to bind and visualize the protein of interest.