Immunohistochemistry (IHC) was performed using the ABC (streptavidin-biotin-peroxidase) method and used the following antibodies: ACE (SC-12187, Santa Cruz Biotechnology, Inc., 1 : 500 dilution) and TGF-β (SC-146, Santa Cruz Biotechnology, Inc., 1 : 500 dilution). Antigen retrieval was performed using citrate buffer pH 6.0 for TGF-β unmasking. The blocking of endogenous peroxidase was done with 3% H2O2 solution and incubation of the slices for 30 minutes at room temperature. Histological sections of the aorta were incubated at 4°C with primary antibodies diluted in blocking solution (3% bovine serum albumin in PBS) for a minimum of 18 hours (overnight). Afterwards, incubation was performed with secondary antibody conjugated with biotin and streptavidin conjugated with horseradish peroxidase (HRP). Then, 3,3-diaminobenzidine (DAB) was used as chromogen. Counterstaining was performed with hematoxylin. Negative control reactions were prepared omitting the primary antibody in one slice for each aorta.
Sc 12187
Manufactured by Santa Cruz Biotechnology
Sourced in United States
SC-12187 is a laboratory equipment product offered by Santa Cruz Biotechnology. It serves as a general-purpose device for various applications in scientific research and analysis. The core function of this equipment is to provide a controlled environment for conducting experiments or procedures. However, a detailed description cannot be provided while maintaining an unbiased and purely factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Sc 146, by Santa Cruz Biotechnology (1 mentions)
Tgf β, by Santa Cruz Biotechnology (1 mentions)
Bbk 120, by ScyTek Laboratories (1 mentions)
Super block, by ScyTek Laboratories (1 mentions)
Camedia 5050z digital camera, by Olympus (1 mentions)
Microphot sa microscope, by Nikon (1 mentions)
A2547, by Merck Group (1 mentions)
2 protocols using sc 12187
1
Immunohistochemical Analysis of Aortic ACE and TGF-β
2
Immunohistochemical Analysis of ACE and SMA
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Sections were deparaffinised, then subjected to antigenic detection of the epitopes with citrate buffer, and an endogenous peroxidase blockade was made with hydrogen peroxidase (3%), a kit (ref: No. BBK 120, Scy Tek, USA) was used to block the endogenous biotin аnd a reagent to block non-specific binding (Superblock, Scy Tek), followed by incubation for 24 hours at 4°C with anti-goat АСЕ -1:300 (sc-12187, Santa Cruz Biotechnology Inc. USA) and monoclonal anti-α smooth muscle actin (A-2547, Sigma) 1:5000 -NRS/TBS/BSA, next incubated with secondary antibody: biotinylated anti-goat (No. AGL015 Scy Tek., USA) for 10 min. The reaction was visualized with 3,3΄-diaminobenzidine tetrachloride (DAB, ScyTek Lab. Inc., USA); counterstaining was performed with Mayer's hematoxylin. As negative controls, sections in which the primary antibodies were replaced by a buffer solution (PBS) were used. Microphotographs were performed with Nikon Microphot SA microscope (Japan), combined with Camedia-5050Z digital camera (Olympus, Japan) at ×100 and ×400 magnification.
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