Table of Contents
Which type of blocking reagent is used for IHC?
In general, serum (same species as the secondary antibody) or bovine serum albumin (BSA) is used for blocking.
What are blocking reagents?
The Blocking Reagent is used to decrease the background in nonradioactive hybridization and detection of nucleic acid hybrids: 1) Blocking prior to the detection procedure is optional; either use the blocking reagent or the appropriate serum from the animal (e.g. sheep) from which the antibody was obtained.
Is blocking necessary for IHC?
Before using specific antibodies to detect antigens by immunohistochemistry (IHC), all potential nonspecific binding sites in the tissue sample must be blocked to prevent nonspecific antibody binding.
Why is BSA used as a blocking agent?
Abstract. BSA blocking is a routine practice among clinicians and researchers working on immunoassays throughout the world. The primary role of BSA is to prevent the non-specific binding by blocking the leftover spaces over solid surface after immobilization of a capture biomolecule.
Why is BSA used for blocking?
BSA blocking is a routine practice among clinicians and researchers working on immunoassays throughout the world. The primary role of BSA is to prevent the non-specific binding by blocking the leftover spaces over solid surface after immobilization of a capture biomolecule.
What is BSA blocking?
What is Bradford reagent used for?
Bio-Rad’s Bradford assays provide a simple and accurate method for determining protein concentrations. The binding of the Bradford reagent to proteins results in a color change which is measured with a spectrophotometer or a microplate reader.
What is blocking in immunohistochemistry?
What is blocking in Immunohistochemistry? Blocking is essential for preventing non-specific binding of antibodies or other reagents to the tissue. Even if the antibody has high specificity towards the target, intermolecular forces can promote non-specific binding to other molecules.
What can be used to block non-specific antibodies in IHC?
View our normal goat serum, normal donkey serum or a complete list of sera for blocking in IHC. Proteins such as BSA or casein may also be used to block non-specific antibody binding, and with these there is no need to match the reagent to the species of the secondary antibody.
What is a protein blocking reagent?
Blocking with sera or a protein blocking reagent prevents non-specific binding of antibodies to tissue or to Fc receptors. Theoretically, any protein that does not bind to the target antigen can be used for blocking.
What is immunohistochemistry (IHC)?
Immunohistochemistry (IHC) experiments often contain one or more blocking steps to reduce background signal and false positives. These steps include Protein blocking Blocking with sera or a protein blocking reagent prevents non-specific binding of antibodies to tissue or to Fc receptors.