{"product_id":"abcam-ab64212","title":"Abcam, ab64212, Avidin\/Biotin Blocking Kit","description":"\u003cp\u003eSize: 15mL\u003cbr\u003e\nAvidin\/biotin blocking kit (ab64212) blocks and minimizes non-specific staining caused by endogenous avidin, biotin and biotin-binding proteins in tissues when used with biotin-based IHC detection. - Supplied as ready-to-use reagents- no dilution required - Cited in over 35 publications\u003cbr\u003e\nProduct details:\u003cbr\u003e\nAvidin Biotin Blocking Kit ab64212 blocks signal from endogenous avidin, biotin and biotin-binding proteins in tissues when used with biotin-based IHC detection (eg. with ABC IHC detection kits).\u003cbr\u003e\nWhen using the kit, firstly an excess of avidin is added to the sample to bind endogenous biotin, that avidin is then blocked with an excess of biotin. Excess biotin and avidin is washed away.\u003cbr\u003e\nThe kit is often used with cells and tissues containing high levels of biotin. This can be indicated by blocking sections with hydrogen peroxide, and then incubating sections with streptavidin-HRP and then DAB; brown DAB staining indicates endogenous biotin. Kidney, liver, spleen especially contain high levels of biotin.\u003cbr\u003e\nThis kit was previously called Endogenous Avidin\/Biotin Blocking Kit.\u003cbr\u003e\nIHC protocol suitable for use with Avidin Biotin Blocking Kit ab64212:\u003cbr\u003e\nFor frozen sections, skip steps 1 and 2.\u003cbr\u003e\n1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.\u003cbr\u003e\n2. Use appropriate\u003cbr\u003e\nantigen retrieval buffer or enzyme\u003cbr\u003e\n(primary antibody dependent) to treat sections. Wash 3 times in buffer.\u003cbr\u003e\n3. Add enough\u003cbr\u003e\nhydrogen peroxide blocking solution\u003cbr\u003e\nto cover the sections. Incubate for 10 minutes. Wash 2 times in buffer.\u003cbr\u003e\nIf necessary, block for endogenous biotin by incubating with avidin block for 15 mins, washing twice, incubating with biotin block for 15 mins, and washing twice.\u003cbr\u003e\n4. Apply\u003cbr\u003e\nprotein block\u003cbr\u003e\nnormal serum\u003cbr\u003e\nfrom same species as secondary antibody) and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash once in buffer.\u003cbr\u003e\n5. Apply primary antibody in\u003cbr\u003e\nantibody diluent\u003cbr\u003e\nand incubate.\u003cbr\u003e\n6. Wash 4 times in buffer. Incubate slide with\u003cbr\u003e\nbiotinylated secondary antibody\u003cbr\u003e\nHRP polymer secondary antibody\u003cbr\u003e\nand skip step 7). Wash 4 times in buffer.\u003cbr\u003e\n7. Apply\u003cbr\u003e\nstreptavidin-HRP\u003cbr\u003e\nand incubate for 10 minutes at room temperature.\u003cbr\u003e\n8. Rinse 4 times in buffer. Place slide in\u003cbr\u003e\nDAB substrate\u003cbr\u003e\nAEC Substrate\u003cbr\u003e\nand incubate until desired color is achieved (1-10 mins). Rinse 4 times in buffer.\u003cbr\u003e\n9. Add enough drops of\u003cbr\u003e\nhematoxylin\u003cbr\u003e\nto cover the section. Incubate for 1 minute.\u003cbr\u003e\n10. Rinse 7-8 times in tap water. Add\u003cbr\u003e\nmounting medium\u003cbr\u003e\nto cover the section.\u003cbr\u003e\nFind complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the\u003cbr\u003e\nIHC kits and reagents guide\u003c\/p\u003e\n\n\u003cp\u003eProperties and Storage Information:\u003cbr\u003e\nShipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C\u003c\/p\u003e","brand":"Abcam","offers":[{"title":"Default Title","offer_id":46847592497321,"sku":"ab64212","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/ar\/products\/abcam-ab64212","provider":"Iright","version":"1.0","type":"link"}