Avidin/Biotin blocking kit
|Product Description||Avidin/Biotin blocking kit|
|Background||Some tissue sections contain endogenous biotin, biotin binding proteins, lectins or non-specific binding substances. Tissues may also bind avidin, biotin, peroxidase biotin, peroxidase streptavidin in avidin biotin based Immunohistochemistry. These tissues will give high background in the absence of biotinylated secondary antibodies. It may be necessary to block the tissue with avidin, followed by biotin. These reagents are added before the primary antibody.
Ready-to-use Reagent A (Avidin) - 15ml
Ready-to-use Reagent B (Biotin) - 15ml
|Applications||ICC/IF, IHC, WB|
|Application Note||IHC/ICC procedure for frozen sections, paraffin sections, cell smears.
1. Deparafinize and hydrate tissue sections through xylene or other clearing agents and graded alcohols.(For frozen sections or cell smears; use unfixed, acetone fixed or appropriate fixative for the antigen in question; for cell smears it may be necessary to permeabilize the cell by detergent, please refer to antibody protocol)
2. Wash 2-3 with distilled or deionized water.
3. Wash slide with Tris/saline buffer, followed by blocking with normal blocking serum, rinse with buffer.
4. Block with reagent A (Avidin) for 5-10 minutes, rinse with buffer.
5. Now block with reagent B (Biotin) for 5-10 minutes, wash thoroughly with buffer.
Note: If antigen retrieval is required it can be applied after this stage.
6. Wash slide with PBS or Tris saline (with 0.02-0.05% nonionic detergent, Triton X100, Tween 20 or NP-40)
7. Follow instructions for IHC/ICC.
1. After protein blocking step, soak blot in avidin reagent A, diluted 1:20 with tris/saline buffer for 5-10 minute.
2. Rinse blot with buffer; followed by soaking in diluted biotin reagent B (diluted 1:20) for 5-10 minutes.
3. Rinse with buffer.
These are guide lines, the optimum incubation times for these reagents and reactions should be determined by the individual lab.
(Please refer to the vial label for the specific concentration)|
|Storage Buffer||10 mM PO4, 200 mM NaCl, pH 7.4, BSA 10 mg/ml and 0.05% Proclin 300|
|Notes||For In vitro laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.|