Adult male and female worms were fixed overnight at 4 °C in 10% neutral buffer formalin. The worms were dehydrated through an ethanol series, therefore infiltrated and embedded in graded paraffin. The embedded worms were cut into 4 µm thick and placed on pre-coated immunohistochemistry slides. Heat-induced antigen retrieval with citrate buffer (pH 6) was used for enhancing tissue antigenicity. EnVision FLEX/HRP (K8002; DAKO, Denmark) and EnVision G/2 System/AP (K5355-11; DAKO, Denmark) kits were used for peroxidase and alkaline phosphatase staining systems, respectively. Subsequence to non-specific binding and endogenous peroxidase blocks, anti-phosphoserine (Merck, USA, AB1603) was applied to the tissue at 1:100 dilution. Regard to the staining systems, the tissue was then incubated in secondary antibody conjugation kits, visualized by either 3, 3-diaminobenzidine (DAB) or liquid permanent red (LPR), and counter stained by hematoxylin. Immunolocalization was examined under a light microscope (BX51, Olympus, Japan) with digital camera (DP20, Olympus, Japan).
Envision g 2 system ap
Manufactured by Agilent Technologies
Sourced in Denmark
The EnVision G/2 System/AP is a microplate reader designed for high-throughput screening and detection of various assays. It provides a flexible platform for fluorescence, luminescence, and absorbance measurements. The system is capable of performing multiple detection modes and supports a wide range of microplate formats.
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Lab products found in correlation
2 protocols using envision g 2 system ap
1
Immunohistochemical Localization of Phosphoserine
2
Immunohistochemical Detection of Macrophages
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For immunohistochemistry, 3µm-thick sections were obtained from para n blocks and mounted on poly-L-lysinecoated slides. Sections were depara nized and rehydrated and subsequently submitted to heat-induced epitope/antigen retrieval for 30 seconds at 125 o C using the Trilogy antigen retrieval buffer (Cell Marque Corporation, Rocklin, CA) in a pressure chamber (Pascal Pressure Chamber, Dako, Carpinteria, CA). Proteinase K antigen retrieval was used when suitable, with subsequent incubation of tissue sections with a mouse anti-macrophage antibody (clone MAC387, 1:400, AbD Serotec, Oxford, UK). Canine lymph node tissue was used as positive control for reactions while negative controls were obtained by using isotype-matched primary antibodies. For anti-macrophage immunohistochemistry, the staining was revealed by using a polymer based detection system (Envision G2 System/AP, rabbit/mouse, Dako) with liquid permanent red as chromogen (Permanent red substrate-chromogen, Dako) and visualized under light microscopy.
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