Deparaffinized and rehydrated formalin fixed samples tumor samples were subjected to antigen retrieval using Dako S1699 Retrieval Buffer at pH 6.1 at 90oC for 30 mins. After a 5 min peroxidase blockade, slides were blocked for 10 min at RT with PBS containing 0.5% BSA, 5% goat serum, and 1X mouse/human FcR blocker (Miltenyi Biotec 130–092-575). Slides were incubated with primary anti-Bnip3 (EPR4034, Abcam) at 1:100 at RT for 30 min, rabbit mAb to cleaved Caspase-3 (Asp175) (5A1E) (Cell Signaling Technology) or anti-Ki67 (SP6) (Thermo-Scientific) at 1:100 overnight at 4oC. IHC was visualized by DAB staining kit (TL-015-HD by Invitrogen), counterstained with hematoxylin, and mounted in non-aqueous solution (Richard-Allen Scientific 4112).
S1699 retrieval buffer
Manufactured by Agilent Technologies
The S1699 Retrieval Buffer is a laboratory reagent designed for use in molecular biology and immunohistochemistry applications. Its core function is to facilitate the retrieval and exposure of target antigens or molecules from fixed tissue samples, enabling further analysis and detection.
Automatically generated - may contain errors
2 protocols using s1699 retrieval buffer
1
Immunohistochemistry of Apoptosis Markers
2
Immunohistochemistry of Apoptosis Markers
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