Formalin-fixed paraffin-embedded (FFPE) human tissues were received mounted on adhesive slides. The slides were baked overnight in an oven at 55°C (Robbin Scientific, Model 1000) and an additional 30 minutes at 65°C (Clinical Scientific Equipment, NO. 100). Tissues were deparaffinized with xylene and rehydrated with graded ethanol baths. Two step antigen retrieval was performed in the Decloaking Chamber (Biocare Medical) using the following settings: set point 1 (SP1), 125°C, 30 seconds; SP2: 90°C, 30 seconds; SP limit: 10°C. Slides were further incubated in hot Target Retrieval Solution, pH 9 (Agilent, S236784–2) for 15 minutes. Slides were then washed in two brief changes of diH2O (~2 seconds) and once for 5 minutes in 1x phosphate buffered saline (PBS), pH 7.4 (Fisher, BP39920). Sections were blocked in 10% normal goat serum (NGS, Vector S-1000), 1% bovine serum albumin (BSA, Sigma A7906) in PBS for 30 minutes at 20°C in a humid chamber, followed by PBS washes. Primary antibodies were diluted in 5% NGS, 1% BSA in 1x PBS and applied overnight at 4°C in a humid chamber, covered with plastic coverslips (Bio-Rad, SLF0601). Following overnight incubation, tissues were washed 3 × 10 min in 1x PBS. Coverslips (Corning; 2980–243 or 2980–245) were mounted in Slowfade Gold plus DAPI mounting media (Life Technologies, S36938).
Bp39920
Manufactured by Thermo Fisher Scientific
The BP39920 is a laboratory equipment product from Thermo Fisher Scientific. It is a compact and portable device designed for performing basic laboratory tasks. The core function of this product is to provide a reliable and consistent platform for common laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bp39920
1
Immunohistochemistry in FFPE Tissue
2
Immunofluorescence Tissue Preparation and IHC
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Tissue preparation and IHC were conducted as previously described (Hammond et al., 2015; Zhang et al., 2020; Zhang et al., 2018b) . Mice were perfused with ice-cold PBS (#BP399-20, Fisher Chemical), and post-fixed in fresh prepared 4% paraformaldehyde (PFA, #1570-S, Electron Microscopy Science, PA) at room temperature for 2 hours. Tissues were washed three times (15 min per each time) using ice-cold PBS and cryoprotected using 30% sucrose (#S5-3, Fisher Chemical) in PBS at 4°C overnight followed by embedding in OCT prior. Twelve microns thick sections were serially collected and stored in -80 °C. For immunohistochemistry, slice was air dry in room temperature for at least 1 hour, and blocked with 10% Donkey serum in 0.1% Triton X-100/PBS (v/v) for 1 hour at room temperature. Tissue was incubated with primary antibodies overnight at 4°C, washed three times in PBST (PBS with 0.1% Tween-20), and incubated with secondary antibodies at room temperature for 2 hours. DAPI was applied as nuclear counterstain. All
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