Paraffin-embedded tissue sections of 4-μm thickness, were deparaffinized in xylene followed by hydration in graded ethanol solution and heated in citrate buffer (pH 6.0) for 5 min. Sections were then blocked with 5% bovine serum albumin (BSA) in Tris-buffered saline (TBS) for 2 h. Slides were then kept overnight at 4 °C with one of the following specific primary antibodies: COX-2 rabbit polyclonal antibody (cat#PA5–23638, Thermo Fischer Scientific), iNOS rabbit polyclonal antibody (cat#RB-9242-P, Thermo Fisher Scientific). Slides were washed with TBS and incubated for 15–20 min with the corresponding secondary antibody. Then, horseradish-peroxidase-conjugated streptavidin solution was added and incubated at room temperature for 15–20 min. The slides were then rinsed with TBS and incubated for 10 min in a solution of 0.02% diaminobenzidine containing 0.01% H2O2. Counter staining was carried out using hematoxylin, and the slides were examined under a light microscope. Immunohistochemical quantification was executed using image analysis software (ImageJ, 1.48a, NIH, USA).
Cox 2 rabbit polyclonal antibody
Manufactured by Thermo Fisher Scientific
Sourced in Canada, United States
The COX-2 rabbit polyclonal antibody is a laboratory reagent used for the detection and study of the cyclooxygenase-2 (COX-2) protein. COX-2 is an enzyme involved in the production of prostaglandins, which play a role in inflammation and other physiological processes. This antibody can be used in various immunoassay techniques, such as Western blotting and immunohistochemistry, to identify and quantify the presence of COX-2 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Inos rabbit polyclonal antibody, by Thermo Fisher Scientific (1 mentions)
Murine il 4, by Thermo Fisher Scientific (1 mentions)
Deae dextran hydrochloride, by Merck Group (1 mentions)
Recombinant human il 1β, by R&D Systems (1 mentions)
Mouse monoclonal β actin antibody, by Cell Signaling Technology (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Wisent (1 mentions)
Phosphate buffered saline (pbs), by Wisent (1 mentions)
3 protocols using cox 2 rabbit polyclonal antibody
1
Immunohistochemical Quantification of COX-2 and iNOS
2
Reagents and Antibodies for Cell Culture
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DMEM, RPMI, fetal bovine serum (FBS) and l -glutamine were purchased from Gibco Life Technologies Ltd., Burlington, Ontario, Canada. Penicillin/streptomycin, phosphate buffered saline (PBS) were from Wisent, St. Bruno, Quebec, Canada. Human recombinant IL-1β was product of R&D Systems (Minneapolis, MN, USA), and LPS endotoxin (Escherichia coli, serotype O128:B12) and DEAE-Dextran hydrochloride of Sigma (Oakville, Ontario, Canada), while murine IL-4 was from Peprotech, Quebec, Canada. Antibodies used in this study were: COX-2 rabbit polyclonal antibody from Thermo Fisher Scientific (Burlington, Ontario, Canada), and COX-2 (C-20) goat polyclonal, NFκB p65 (C-20), p50 (H-119) and Lamin A (H-102) rabbit polyclonal antibodies from Santa Cruz (Dallas, TX, USA). ESE-1 rabbit monoclonal antibody was produced in our laboratory in collaboration with Epitomics, Burlingame, CA, USA [22 (link)]. Hsp90 rabbit polyclonal and β-actin mouse monoclonal antibodies were purchased from Cell Signaling Technology (Whitby, Ontario, Canada).
3
Immunohistochemical Analysis of COX-2
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Isolated hippocampi (n = 3) were fixed in 10% (v/v) formalin for 24 h to perform the immunohistochemistry assay. Paraffin-embedded tissue sections of 3–5 μm thickness were first deparaffinized with xylene and then hydrated in graded ethanol solution and heated in citrate buffer (pH 6.0) for 5 min. Next, the sections were blocked with 5% bovine serum albumin in phosphate buffered saline (PBS) for 2 h. The slides were then incubated overnight at 4 °C with COX-2 rabbit polyclonal antibody (1:100, Thermo Fischer Scientific, Waltham, MA, USA). The slides were rinsed with PBS and incubated for 10 min in a solution of 0.02% diaminobenzidine. Sections were counterstained with haematoxylin, dehydrated, cleared in xylene and then cover slipped for light microscopic examination. Six fields were randomly selected from each section, and positive signals within the section were highlighted, measured, and expressed as the percent area of expression of COX-2 in the immunostained tissue sections using Leica Microsystems (GmbH, Wetzlar, Germany).
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