Different organs were harvested and imbedded in OCT and kept frozen at –80 °C. Frozen sections were cut into 10-μm-thick slices with cryostat. Different slices were fixed with z-fix for 30 min at room temperature and washed with PBS three times. 2% BSA in PBS was used to block the slices at room temperature for 1 h. Anti-PSMA antibody (Abcam ab133579, 1:150, 2% BSA in PBS) was added onto the slices and incubated at 4 °C overnight followed by PBS wash for 3 times (5 min each time). Goat-anti- rabbit secondary antibody (R&D, 1:100, 2% BSA in PBS) was added and slices were incubated for 1 h at room temperature. After washing with PBS three times, the slices were mounted with mounting solution containing DAPI. Fluorescence images were acquired by a fluorescence microscope (Olympus). Ki67, TUNEL, and H&E staining were also performed according to the kit instructions.
Goat anti rabbit secondary antibody
Manufactured by R&D Systems
Sourced in United States
Goat anti-rabbit secondary antibody is a laboratory reagent used in immunoassays to detect and quantify rabbit primary antibodies. It binds to the Fc region of rabbit antibodies, allowing for their visualization or isolation.
Automatically generated - may contain errors
Lab products found in correlation
Anti f4 80 antibody, by Abcam (2 mentions)
Ab133579, by Abcam (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Complete protease inhibitor, by Merck Group (1 mentions)
Anti phospho stat3 tyr705, by Cell Signaling Technology (1 mentions)
Bca protein determination kit, by Thermo Fisher Scientific (1 mentions)
Anti total stat3, by Abcam (1 mentions)
Anti il 6, by Abcam (1 mentions)
4 protocols using goat anti rabbit secondary antibody
1
Immunohistochemical Analysis of PSMA Expression
2
Protein Extraction and Western Blot Analysis
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Proteins from tissue (1 × 2 mm) or BON1 cells were extracted using RIPA buffer containing complete protease inhibitors (Sigma-Aldrich) according to the manufacturer’s instructions, and the protein concentration was determined using a BCA Protein Determination Kit (Thermo Scientific) following the manufacturer’s instructions. Western blots were processed as previously described (31 (link)) using the following antibodies: anti-IL-6, anti-total-STAT3, and anti-ATX (all from Abcam); and anti-phospho-STAT3 (Tyr705) (Cell Signaling Technology). A goat anti-rabbit secondary antibody was used (R&D Systems). Protein expression was quantified using ImageJ (NIH).
3
Immunohistochemical Staining of F4/80
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The prepared para n sections were put into an oven at 60℃ for one hour and then dewaxed in xylene and ethanol with different concentration gradients. After dewaxing, the antigen was repaired followed by incubation with anti F4/80 antibody (dilution: 1:200, Abcam, USA) at 4 ℃ for 12 hours. The slides were washed twice with PBS followed by incubation with goat anti-rabbit secondary antibody (R&D, USA) at 37 ℃ for 1 hour. Again, the slides were washed twice with PBS and the reaction visualized with diaminobenzidine (DAB) for 8 minutes. The reaction was terminated by rinsing with water followed by hematoxylin-eosin staining. Finally, the slide sections were observed under an inverted optical microscope and the results analyzed by Image J software.
4
Immunohistochemical Staining of F4/80
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The prepared para n sections were put into an oven at 60℃ for one hour and then dewaxed in xylene and ethanol with different concentration gradients. After dewaxing, the antigen was repaired followed by incubation with anti F4/80 antibody (dilution: 1:200, Abcam, USA) at 4 ℃ for 12 hours. The slides were washed twice with PBS followed by incubation with goat anti-rabbit secondary antibody (R&D, USA) at 37 ℃ for 1 hour. Again, the slides were washed twice with PBS and the reaction visualized with diaminobenzidine (DAB) for 8 minutes. The reaction was terminated by rinsing with water followed by hematoxylin-eosin staining. Finally, the slide sections were observed under an inverted optical microscope and the results analyzed by Image J software.
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