Description
Product Introduction:1. The Biotin Assay Blocking Kit is a kit primarily designed for blocking tissue and cell samples to significantly reduce background during detection processes such as immunohistochemistry (IHC), immunocytochemistry (ICC), immunofluorescence (IF), and fluorescence in situ hybridization (FISH). These detection methods are based on biotin and streptavidin or avidin, including the SABC (Streptavidin-Biotin Complex) method and ABC (Avidin-Biotin Complex) method.2. This kit contains two types of blocking buffers: Biotin Blocking Buffer (100X): Contains streptavidin and other components, which can fully bind to and block endogenous biotin, and reduce non-specific binding to labeled streptavidin in subsequent steps. Streptavidin/Biotin-Binding Blocking Buffer (100X): Contains biotin, which can fully block endogenous biotin-binding proteins and the streptavidin bound during the previous blocking step. Through this two-step blocking process, the goal of significantly reducing background can be achieved.3. This kit is suitable for blocking frozen sections and paraffin sections, and also applicable for blocking cultured cells.Product Features:1. Easy to Use – Can be performed simultaneously with the blocking step of conventional immunostaining, significantly saving blocking time.2. Stable Performance – Effectively reduces non-specific staining and improves detection sensitivity.Product Components and Storage Conditions:Product numberComponent200TStorageB1209118ABiotin Blocking Buffer (100×)200 µl-20ºCB1209118BStreptavidin/Biotin-Binding Blocking Buffer (100×)200 µl-20ºCOperating Steps:1. For immunostaining assays such as IHC, ICC, and IF:a. Dilution of blocking buffers: Based on the required volume, dilute Biotin Blocking Buffer and Streptavidin/Biotin-Binding Blocking Buffer to 1X using an appropriate conventional blocking buffer.b. After thorough washing of the sections and before primary antibody incubation, add an appropriate amount of Biotin Blocking Buffer and incubate for 10-30 minutes. Incubation for 10 minutes is recommended when diluted with a rapid immunostaining blocking buffer; incubation for 30 minutes is recommended when diluted with a regular blocking buffer.c. Wash the samples with washing buffer 3 times, 5 minutes each.d. Add an appropriate amount of Streptavidin/Biotin-Binding Blocking Buffer and incubate for 10-30 minutes. Incubation for 10 minutes is recommended when using a rapid immunostaining blocking buffer; incubation for 30 minutes is recommended when using a regular blocking buffer.e. Wash the samples with washing buffer 3 times, 5 minutes each.f. Proceed with subsequent steps such as primary antibody incubation according to conventional procedures.2. For in situ hybridization (FISH) detection:a. Dilution of blocking buffers: Based on the required volume, dilute Biotin Blocking Buffer and Streptavidin/Biotin-Binding Blocking Buffer to 1X using an appropriate diluent. The washing buffer for in situ hybridization can be used for dilution.b. Before hybridization with biotin-labeled probes, add an appropriate amount of Biotin Blocking Buffer and incubate for 15-30 minutes.c. Wash the samples with washing buffer 3 times, 5 minutes each.d. Add an appropriate amount of Streptavidin/Biotin-Binding Blocking Buffer and incubate for 15-30 minutes.e. Wash the samples with washing buffer 3 times, 5 minutes each.f. Proceed with subsequent steps such as hybridization with biotin-labeled probes according to conventional in situ hybridization detection procedures.Precautions:1. This kit can also perform the above-mentioned blocking steps after the primary antibody incubation is completed and the sample is washed 3-4 times with washing buffer, followed by subsequent steps such as secondary antibody incubation.2. This kit can simultaneously perform biotin-related blocking and conventional blocking, eliminating the need for additional conventional immunostaining blocking steps.3. For your safety and health, please wear a lab coat and disposable gloves during operation