BMMNCs smears were uniformly spread on poly-lysine coated slides at 2×105 cells/slide, and were fixed with 4% paraformaldehyde at room temperature for 20 min prior to being washed with PBS 3 times. The slides were incubated with 0.3% tritron-100 at room temperature for 30 min to permeabilize the cells. Following washing with PBS for 3 times, the slides were blocked with 10% bovine serum albumin at room temperature for a further 30 min. Slides were then incubated with a rabbit anti-human PRPS1 polyclonal antibody (dilution, 1:200; cat. no. ab137577; Abcam) at 4°C overnight followed by a peroxidase-conjugated goat anti-rabbit secondary antibody (dilution, 1:50; cat. no. TA140003; OriGene Technologies, Inc., Rockville, MD, USA) at room temperature for 30 min. The slides were then stained with diaminobenzidine at room temperature for 2 min and counterstained with hematoxylin at room temperature for 40 sec. Images were acquired using a light microscope with Olympus BX51 system (Olympus Corporation, Tokyo, Japan) at ×100 magnification.
Peroxidase conjugated goat anti rabbit secondary antibody
Manufactured by Abcam
Sourced in United States
Peroxidase-conjugated goat anti-rabbit secondary antibody is a laboratory reagent used to detect and quantify target proteins in various immunoassay techniques. It consists of a goat-derived antibody that binds to rabbit primary antibodies, conjugated with the enzyme peroxidase. This secondary antibody can be used to amplify and visualize the signal from primary antibody-antigen interactions.
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Lab products found in correlation
2 protocols using peroxidase conjugated goat anti rabbit secondary antibody
1
Immunohistochemical Analysis of PRPS1 in BMMNCs
2
Western Blot Analysis of TRPM8
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Mesenteric arteries were removed and prepared as described above. The mesenteric artery was homogenized in ice-cold RIPA lyses buffer containing a protease inhibitor cocktail (Roche, Mannheim, Baden-Württemberg, Germany). The homogenate was centrifuged at 4°C at 10,000 rpm for 10 min, the supernatant was collected, and the protein concentration was calculated using the Lowry method [27 (link)]. The protein samples were denatured and separated on a denaturing SDS/7.5% polyacrylamide gel, then transferred to a nitrocellulose membrane (PerkinElmer Inc, Waltham, Massachusetts, USA). The membrane was blocked with 5% (wt/vol) milk in TBS containing 0.1% Tween 20 (TBST) for 1 h at room temperature, followed by overnight incubation at 4°C with the specific primary rabbit polyclonal antibody against TRPM8 (1:1000 dilution; Abcam, Cambridge, Massachusetts, United States). After washing, the membrane was incubated with peroxidase-conjugated goat-anti-rabbit secondary antibody (1:3000 dilution; Abcam, Cambridge, Massachusetts, United States) at room temperature for 2 h. Protein loading was confirmed and normalized by comparing to β-actin levels.
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