Antibodies – Blocking/Neutralization Methods, Techniques, Protocols
• Definition: A method to test the specificity of antibodies or to reduce nonspecific background staining, used as a negative control, competition control, absorption, isotype control, etc..
• Demonstration of antibodies specific antigen (peptide / protein) blocking – It’s not unusual to see more than one lane in the West, when probed with antibodies or see more diffuse staining in immunolocalization studies. This raises the question of the group (s) / color specific. Examined the specificity of the antibody competition with excess antigen (peptide or protein) or immuno-neutralization antigen.
• A new blocking method for application of murine monoclonal antibody to mouse tissue sections – antigen detection with the primary antibody of the same type as the test tissue is complicated by a high level of background staining using an indirect immunohistochemical methods. This severely limits the use of mouse monoclonal antibodies on mouse tissue, the most widely used experimental model system, there is no method to lock these feelings. Here we show that the background color is in the system generates a large extent through the secondary antibody binding to Fc and Fab immunoglobulin and other endogenous tissuecomponents. A simple and effective strategy to block created using papain digested whole fragments of anti-mouse secondary enriched AGS unlabeled Fc fragment of the IGS.
• Blocking unwanted non-specific staining immunohistochemistry – Source unwanted color,
but poor knowledge of the reactivity of antibodies and malignancy, is associated with endogenous enzyme or fluorochrome, endogenous biotin, endogenous antibody activity (Fc receptor), Crossreactivity secondary reagents with endogenous proteins.
• Sample Preparation – Blocking – When using antibodies to stain specimens is often necessary to block the sample in order to reduce non-specific binding. Non-specific binding can occur for several reasons: The UN reacted aldehydes, may be a link between antibodies to inappropriate structures (especially with glutaraldehyde) or very highly charged hydrophobic structure models can “capture” antibody, or when using polyclonal antibodies, low affinity IgG that can bind speciously structures are not interested in.