Avidin-Biotin Blocking Set
Description:
Avidin-Biotin Endogenous Blocking Reagent Set
Application:
For blocking endogenous avidin and or biotin in tissues in immunohistochemistry.
Presentation:
Tissue containing endogenous biotin such as kidney and brain, can successfully be subjected to immunocytochemical staining procedures utilizing the Avidin Biotin Complex (ABC) technology if pretreated sequentially with avidin followed by biotin. This blocking procedure renders inactive any available biotin which the specimen may possess. It is achieved by first saturating these residues with avidin and later completely inactivating the bound avidin with an excess of biotin. These products are supplied in PBS with 0.05% sodium azide.
Use this procedure only if the presence of interfering biotin is suspected. When using the capillary reagent transport technology such as with the MicroProbe or Code-On stainers, apply the required drops of Reagent A and the Reagent B to the rubber Isolon plates, and then introduce the capillary gaps at their respective times.
Specificity:
Tissue containing endogenous biotin such as kidney and brain, can successfully be subjected to immunocytochemical staining procedures utilizing the Avidin Biotin Complex (ABC) technology if pretreated sequentially with avidin followed by biotin. This blocking procedure renders inactive any available biotin which the specimen may possess. It is achieved by first saturating these residues with avidin and later completely inactivating the bound avidin with an excess of biotin. Use this procedure only if the presence of interfering biotin is suspected.
Storage:
Store at 4C, do not freeze.
Notes:
Use of Avidin-Biotin Endogenous Blocking Reagent Set: If using paraffin embedded tissue sections, first remove the paraffin with xylene followed by graded alcohol. For frozen sections, defrost the specimens and hydrate in 1X Immunoassay Buffer* or phosphate buffered saline, pH 7.5 (PBS). If required, treat the sections for the destruction of endogenous enzymes such as peroxidase or alkaline phosphatase.
Following this treatment, cover the sections with 1X Immunoassay Buffer and allow to stand for 5 minutes.
1. Remove all liquid from the section.
2. Add four drops of Reagent A and incubate for 15 minutes at the same temperature as to be used in the overall procedure.
3. Rinse sections thoroughly with 1X Immunoassay Buffer.
4. Apply four drops of Reagent B and incubate for 15 minutes.
5. Rinse specimens thoroughly with 1X Immunoassay Buffer.
6. Proceed with the staining protocol as usual.
If background noise persists after this treatment, it is unlikely to be due to endogenous biotin content.
* Whenever 1X Immunoassay Buffer is suggested, you may substitute with 1X Automation Buffer (Cat. No. M30).