Tissue sections were subjected to immunohistochemistry for the detection of the androgen receptor (AR), estrogen receptor-alpha (ER-α), and proliferating cell nuclear antigen (PCNA). Primary antibodies reactive to AR (rabbit polyclonal IgG, N-20, Santa Cruz Biotechnology, CA, USA), ER-α (rabbit polyclonal IgG, MC-20, Santa Cruz Biotechnology), p63 (mouse monoclonal IgG2a, sc-843, 4A4, Santa Cruz Biotechnology, CA, USA) and PCNA (mouse monoclonal IgG2a, SC 56, Santa Cruz Biotechnology, CA, USA) were employed at a dilution of 1:100. Peroxidase-conjugated specific antibodies (Sigma Chemical Co., Saint Louis, MO, USA) or polymers (Post Primary Block and polymer, Novocastra, Newcastle Upon Tyne, UK; DAKO Envision™ + Dual link system-HRP, K4061) were used as secondary antibodies and incubated with samples for 45 min at 37°C. The sections were reacted with diaminobenzidine and counterstained with Harris's hematoxylin. The histological sections were analyzed with a Olympus BX60 light microscope (Olympus, Tokyo, Japan).
Post primary block and polymer
Manufactured by Leica Biosystems
Sourced in United Kingdom
The Post Primary Block and Polymer is a laboratory equipment product from Leica Biosystems. It is used to enhance the visualization of immunohistochemical staining in tissue samples. The product functions to amplify the signal of the primary antibody, enabling clearer and more distinct staining results.
Automatically generated - may contain errors
3 protocols using post primary block and polymer
1
Immunohistochemical Analysis of Prostate Markers
2
Immunohistochemical Analysis of AR, ERα, and PCNA
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Tissue sections were subjected to immunohistochemistry for the detection of the androgen receptor (AR), as described in protocols applied to the prostate (Cordeiro et al. 2008) , oestrogen receptor-alpha (ERα) and PCNA. Primary antibodies reactive to AR (rabbit polyclonal IgG, N-20, sc-816, Santa Cruz Biotechnology, Santa Cruz, CA, USA), ERα (rabbit polyclonal IgG, MC-20, sc-542, Santa Cruz Biotechnology) and PCNA (mouse monoclonal IgG 2a , SC 56, Santa Cruz Biotechnology) were employed at a dilution of 1:100. Polymers (Post Primary Block and Polymer, Novocastra™, RE7260-K, Newcastle Upon Tyne, UK; DAKO Envision tm + Dual link system-HRP, K4061; DAKO, North America, Inc., Carpinteria, CA, USA) were used as secondary antibodies according to the procedures described by the manufacturers. The sections were stained with diaminobenzidine and counterstained with Harris's haematoxylin. The histological sections were analysed using an Olympus BX60 light microscope (Olympus).
3
Immunohistochemical Analysis of Prostate Markers
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Prostate sections were subjected to immunohistochemistry (n = 3 animals per group) for the detection of the androgen receptor (AR), as described in protocols applied to the prostate (Cordeiro et al. 2008) , oestrogen receptor-alpha (ERα), α-actin and PCNA. Primary antibodies reactive to AR (rabbit polyclonal IgG, N-20, sc-816; Santa Cruz Biotechnology, Santa Cruz, CA, USA), ERα (rabbit polyclonal IgG, MC-20, sc-542; Santa Cruz Biotechnology), α-actin (mouse monoclonal IgG2a, sc-32251, IA4; Santa Cruz Biotechnology) and PCNA (mouse monoclonal IgG 2a , SC 56; Santa Cruz Biotechnology) were employed at a dilution of 1:100. Polymers (Post Primary Block and Polymer, Novocastra™, RE7260-K, Newcastle Upon Tyne, UK; DAKO Envision™ + Dual Link System-HRP, K4061; DAKO, North America, Inc., Carpinteria, CA, USA) were used as secondary antibodies according to the procedures described by the manufacturers. Negative control of reaction was performed by omitting the primary antibody. The sections were stained with diaminobenzidine and counterstained with Harris's haematoxylin. The histological sections were analysed using an Olympus BX60 light microscope (Olympus).
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