Prostate ventral lobes and the bladder were harvested for histological analyses after cystometry. One part of the prostate and the bladder was fixed in buffered 10% formaldehyde solution for 24 hr, embedded in paraffin, cut with a microtome, and stained with hematoxylin-eosin for evaluating tissue inflammation. The remainder of the prostate was used for immunohistochemical staining. Paraffin embedded prostate sections were placed on silicone-coated slides. After deparaffinization in xylene and rehydration using graded alcohol solutions, the sections were fixed with 10 mM sodium citrate pH 6 at 105°C by Autoclave (TOMY SEIKO co. Ltd.) for antigen retrieval. The tissues were rinsed with phosphate buffered saline (PBS), transferred to 0.3% hydrogen peroxide for 10 min to block peroxidase activity, and rinsed with distilled water and PBS. The sections were blocked with 10% normal goat serum (NICHIREI CORPORATION.) for 30 min. These tissues were incubated with an anti-ERβ antibody (1:1,000; Santa Cruz Biotechnology, Inc.) for 1 hr at room temperature. After washing with PBS, the tissue sections were incubated for 30 min with HRP-labeled polymer anti-rabbit antibody (DAKO.) at room temperature. After washing with PBS, the color was developed using the Dako Cytomation Liquid DAB Substrate Chromogen System (DAKO). The tissues were counterstained with hematoxylin.
Hrp labeled polymer anti rabbit antibody
Manufactured by Agilent Technologies
Sourced in United States, Denmark
The HRP-labeled polymer anti-rabbit antibody is a detection reagent used for immunohistochemistry and Western blotting applications. It consists of a polymer backbone with multiple horseradish peroxidase (HRP) enzymes and anti-rabbit antibodies. This product provides signal amplification and can be used to detect target proteins recognized by rabbit primary antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Dakocytomation liquid dab substrate chromogen system, by Agilent Technologies (1 mentions)
Anti erβ antibody, by Santa Cruz Biotechnology (1 mentions)
Autoclave, by TOMY (1 mentions)
Envision system, by Agilent Technologies (1 mentions)
Bz x810, by Keyence (1 mentions)
Ab182981, by Abcam (1 mentions)
Rabbit anti mouse cd68, by Wuhan Servicebio Technology (1 mentions)
3 3 diaminobenzidine substrate kit, by Boster Bio (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Pannoramic midi slide scanner, by 3DHISTECH (1 mentions)
Normal goat serum (ngs), by Thermo Fisher Scientific (1 mentions)
Cox 2, by Abcam (1 mentions)
Abc reagent, by Agilent Technologies (1 mentions)
Interleukin 6 (il 6), by Abcam (1 mentions)
Tgf β1, by Abcam (1 mentions)
4 protocols using hrp labeled polymer anti rabbit antibody
1
Histological Analysis of Prostate and Bladder
2
Histological and Immunohistochemical Analysis of Lung Tissue
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The extracted lungs were fixed in 4% paraformaldehyde and embedded into paraffin, then were cut into 5-μm-thick sections. Hematoxylin-eosin (HE) staining was performed with hematoxylin for 5 min and eosin for 5 min. Masson's trichrome staining was performed using the standard Masson's trichrome stain kit protocol (HT15, Sigma Aldrich).
For immunohistochemistry, antigen retrieval was performed by 10 mmol/L of Tris-HCl (pH 8.8) buffer. Following the inhibition of endogenous peroxidase using 3% H2O2 in Tris-buffered saline (TBS), the sections were incubated overnight at 4 °C with a rabbit monoclonal anti-ACSL4 antibody and a rat monoclonal anti-Ly6G antibody (1:3000, 1:500 dilution each). The sections were washed three times with TBS, and then the corresponding secondary antibodies were applied and incubated for 1 h at room temperature. Immunohistochemistry was performed using a Dako EnVision + System with an HRP-labeled polymer anti-rabbit antibody (DAKO, Carpinteria, CA, USA) according to the manufacturer's instructions and then counterstained with hematoxylin. The image was captured with a microscope (BZ-X810; Keyence, Osaka, Japan).
For immunohistochemistry, antigen retrieval was performed by 10 mmol/L of Tris-HCl (pH 8.8) buffer. Following the inhibition of endogenous peroxidase using 3% H2O2 in Tris-buffered saline (TBS), the sections were incubated overnight at 4 °C with a rabbit monoclonal anti-ACSL4 antibody and a rat monoclonal anti-Ly6G antibody (1:3000, 1:500 dilution each). The sections were washed three times with TBS, and then the corresponding secondary antibodies were applied and incubated for 1 h at room temperature. Immunohistochemistry was performed using a Dako EnVision + System with an HRP-labeled polymer anti-rabbit antibody (DAKO, Carpinteria, CA, USA) according to the manufacturer's instructions and then counterstained with hematoxylin. The image was captured with a microscope (BZ-X810; Keyence, Osaka, Japan).
3
Histological Analysis of Skeletal Muscle
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For histological analysis, the gastrocnemius muscles were fixed in 4% paraformaldehyde for 24 h, embedded in paraffin, cut into sections 5 μm thick, and stained with hematoxylin–eosin (HE). The percentage of tissue necrosis was quantified using Image Pro Plus 6.0 software (Media Cybernetics, Silver Spring, MD, USA). For the evaluation of the capillary density, the specimens were incubated in rabbit monoclonal anti-CD31 antibody (ab182981; Abcam, Cambridge, UK) overnight at 4 °C. HRP-labeled polymer anti-rabbit antibody (Dako, Glostrup, Denmark) was then added for 30 min at 37 °C. A 3,3′-diaminobenzidine substrate kit (Boster, Wuhan, China) was used to visualize the reaction. For the detection of infiltrated monocytes/macrophages, sections were stained using rabbit anti-mouse CD68 (Servicebio, Wuhan, China). Whole slide images were obtained using a Pannoramic MIDI slide scanner (3DHISTECH Kft., Budapest, Hungary). The capillary density was calculated as the total number of vessels identified in the total section area. The level of monocytes/macrophages infiltration was defined as CD68+ cells/mm2. Three sections from each specimen were used for analysis.
4
Immunohistochemical Analysis of Prostate Samples
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