The cryosections of the prostate tissue were washed with PBS for 10 min to remove the OCT compound and then fixed with anhydrous methanol at −20 °C for 15 min in the dark. Then, the cryosections were washed three times with PBS. The cryosections were dried, and the tissue to be stained was circled by the PAP pen (ab2601, Abcam). Then, the samples were blocked with 200 μL anti-goat serum at room temperature for 60 min. Afterward, the anti-goat serum on the slice surface was discarded, and 100 μL of Cy-KUE-OA (5 μmol/L) and Alexa Fluor® 488-conjugated mouse monoclonal anti-PSMA antibody (ab187570, Abcam) were added into each well, and incubated at 4 °C overnight in the dark. The next day, the cryosections were washed three times with PBS for 10 min each. After drying the samples, 10 μL of DAPI anti-fluorescence quenching agent was added, and then the samples were examined using a Leica TCS SP8 X Confocal Microscope.
Ab2601
Manufactured by Abcam
Ab2601 is a laboratory equipment product. It is designed for use in scientific research and experiments. The core function of this product is to [insert factual and concise description of the product's core function without any extrapolation or interpretation].
Automatically generated - may contain errors
Lab products found in correlation
Tcs sp8 x confocal microscope, by Leica (1 mentions)
G0002, by Wuhan Servicebio Technology (1 mentions)
Gc305006, by Wuhan Servicebio Technology (1 mentions)
Confocal laser scanning microscope, by Leica (1 mentions)
Citrate antigen retrieval solution, by Solarbio (1 mentions)
G1401, by Wuhan Servicebio Technology (1 mentions)
C1002, by Beyotime (1 mentions)
2 protocols using ab2601
1
Prostate Tissue Staining and Analysis
2
Immunofluorescent Detection of UCP1 in Paraffinized Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All tissues were fixed in 4% paraformaldehyde over 24 h at 4°C, then dehydrated and embedded into paraffin.
After that, tissues were sectioned into thick slices (4 to 6 µm) and stained with hematoxyline and eosin (H&E).
For immunofluorescence, slices were first boiled for 15min in Citrate Antigen Retrieval solution (C1032; Solarbio) diluted with double distilled water to pH 6.0. After cooling to room temperature, we circled the tissue with a PAP pen (ab2601; Abcam), dropped 3% bovine serum albumin (GC305006; Servicebio) in PBS (G0002; Servicebio), which covered the tissue and it then got blocked for 30min. The slices were incubated with rabbit polyclonal anti-UCP1 primary antibody (23673-1-AP; Proteintech) 1:50 diluted in PBS overnight at 4°C. The slices were rinsed in PBS for 3 times, followed by treating with Goat Anti-Rabbit IgG H&L/Alexa Fluor 594 (bs-0295G-AF594; Bioss) incubated at room temperature for 50 min. Every move since this step needs to avoid light. DAPI (C1002; Beyotime) stained uncleus for 10 min and the slices were washed in PBS for 3 times and sealed with anti-fade mounting medium (G1401; Servicebio) for longer storage. Photomicrographs were taken under a confocal laser-scanning microscope (Leica; Germany). Images shown are representative results of at least three biological replicates.
After that, tissues were sectioned into thick slices (4 to 6 µm) and stained with hematoxyline and eosin (H&E).
For immunofluorescence, slices were first boiled for 15min in Citrate Antigen Retrieval solution (C1032; Solarbio) diluted with double distilled water to pH 6.0. After cooling to room temperature, we circled the tissue with a PAP pen (ab2601; Abcam), dropped 3% bovine serum albumin (GC305006; Servicebio) in PBS (G0002; Servicebio), which covered the tissue and it then got blocked for 30min. The slices were incubated with rabbit polyclonal anti-UCP1 primary antibody (23673-1-AP; Proteintech) 1:50 diluted in PBS overnight at 4°C. The slices were rinsed in PBS for 3 times, followed by treating with Goat Anti-Rabbit IgG H&L/Alexa Fluor 594 (bs-0295G-AF594; Bioss) incubated at room temperature for 50 min. Every move since this step needs to avoid light. DAPI (C1002; Beyotime) stained uncleus for 10 min and the slices were washed in PBS for 3 times and sealed with anti-fade mounting medium (G1401; Servicebio) for longer storage. Photomicrographs were taken under a confocal laser-scanning microscope (Leica; Germany). Images shown are representative results of at least three biological replicates.
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