With the broadest range of antibody generation platforms, Creative Biolabs offers high performance antibodies and outstanding customer services. As a well known expert in the field of antibody, our talented scientists have built expertise and experience to advance and accelerate your research, diagnosis or drug discovery programs based on antibody. Now we display an overview of application methods and techniques for use with your antibody.
ELISA assays are one of the most commonly employed screening methods based upon the principle of antibody/antigen binding. They are widely used in quantification and characterization of specific analytes or molecular interactions. Theoretically, antibodies against the target of interest are linked to a reporter enzyme. The enzyme can catalyze the production of a colorimetric molecule once its substrate is added. Thus, the final reaction produces a detectable signal, most commonly a color change. This reaction can be measured using a spectrophotometer and is representative of antigen concentration within a sample.
ELISA has different formats and the most appropriate format for each experiment often depends on many factors, such as the desired sensitivity, specificity and assay time. The relevant information about ELISA assays is as follows:
Western blots (also known as western blotting or protein immunoblot) are often used to determine relative protein levels between samples. The analytical technique also is used to establish the molecular weight of the target, which may endow insight into its post-translational processing.
Commonly, protein samples from tissue or cell lysates are separated by gel electrophoresis based on their different molecular weight. Subsequently, separation samples are transferred to a membrane, where antibodies are applied to probe the protein of interest through various methods such as staining, immunofluorescence, and radioactivity. Related information is as follows:
IHC represents one of the most common assays for immunostaining to study tissue-specific and subcellular localization of an antigen within a sample. The method shows the advantage of characterizing protein expression in the context of intact tissue compared with Western blot or ELISA.
In theory, IHC staining is performed using antibodies that can specifically recognize and bind to antigens or antibody-antigen complexes in biological tissues. Thus, the target protein can be visualized through chromogenic, radioactive or fluorescent substrates. Besides, various techniques can be used for sample preparation and visualization, and the suitable method used should depend on the type of sample under investigation and the degree of sensitivity required. Related information about IHC assays is as follows:
FC is a technique used in the measurement of certain chemical and physical properties of cells or particles. Parameters detection include cell-size and the expression of cell-surface and intracellular markers.
FC measures the fluorescence emitted by labeled antibodies that bind individual cells in a mixed population. Fluorescence-activated cell sorting (FACS) is a sophisticated system, which quantifies the fluorescent signal and separates the cells from a mixed population that contains preselected characteristics (such as the fluorescence intensity, size and viability). FC can be used in basic research, clinical practice, and clinical trials.
Immunoprecipitation is a useful and versatile technique that is widely used in the isolation and purification of individual and complexed proteins. Simply, antibodies are primarily immobilized on solid-phase substrates (such as magnetic/agarose beads), which capture antigens from complex solutions. For chromatin immunoprecipitation (ChIP), it is a procedure used to determine whether a given protein binds to a specific DNA sequence in vivo.
Related information about IP assays is as follows:
ELISPOT is a method based on ELISA used for the detection of secreted proteins of cells, such as cytokines and growth factors. The technique is used for quantification and comparison of immune responses to various stimuli.
In short, cells are cultured in 96-well plates with antibody-coated PVDF or nitrocellulose membranes, and the secreted protein of interest binds the antibody. The interactions between the antibody and antigen are detected using a secondary antibody, and the protein is visualized as a spot color under the parent cell. In addition, the membranes can be scanned and analyzed to quantify the number/percentage of cells secreting the protein.
Creative Biolabs is dedicated to providing the best antibody products and services with the highest level of quality. With years of know-how, our experienced scientists will work closely with you to provide help. We are a reliable partner that can help you with the greatest chance to succeed. If you have any special requirements in antibody methods or techniques, please feel free to contact us. We are looking forward to working together with your attractive projects.
