Dorsal skin samples were embedded in OCT and sectioned at 8 µm thickness. Sections were fixed in formalin for 10 min, followed by two H2O washes (10 min each). Sections were blocked with blocking buffer (10% donkey serum in 1xPBST) for 1 h. Primary antibodies were diluted in blocking buffer and applied to the tissue and incubated overnight at 4 °C. Secondary antibodies were diluted in 1xPBST for 1 h at room temperature following three 1xPBST washes (5 min). Sections were washed in 1xPBST three times (5 min) and mounted with Fluoroshield with DAPI (Abcam, ab104139). Images were taken using the Leica DM7200 fluorescence imaging platform with LAS X version 3.7.5. Antibodies used: F4/80 1:600 (Biolegend, Cat#123101); Ly6G 1:600 (Biolegend, Cat#127601); CD3 1:800 (Biolegend, Cat#100201); Cox-2 1:600 (Cayman, Cat#160106); Ptges2 1:200 (Abclonal, Cat#A7137); Ptges3 1:200 (Abclonal, Cat#A5194); Ptger2 1:500 (Abcam, Cat#ab167171); Cd49f 1:100 (BD Biosciences, Cat#555734); Dct 1:600 (Abcam, Cat#ab221144), together with suitable Alex Fluor secondary antibodies (Abcam, Cat#ab150072, ab150149, ab150152, Fisher, Cat#A21207)
Ptger2
Manufactured by Abcam
Sourced in United States
Ptger2 is a laboratory equipment product. It serves as a core component in various research applications. The detailed specifications and intended use of this product are not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Cox 2, by Cayman Chemical (2 mentions)
Dm7200, by Leica camera (2 mentions)
Ptges2, by ABclonal (2 mentions)
Ptges3, by ABclonal (2 mentions)
Cd49f, by BD (2 mentions)
F4 80, by BioLegend (2 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Wuhan Servicebio Technology (1 mentions)
β actin, by Wuhan Servicebio Technology (1 mentions)
Col 1, by Wuhan Servicebio Technology (1 mentions)
α sma, by Wuhan Servicebio Technology (1 mentions)
Total protein extraction kit, by Keygen Biotech (1 mentions)
Bca protein assay kit, by Keygen Biotech (1 mentions)
3 protocols using ptger2
1
Multiparametric Immunofluorescence Imaging
2
Western Blot Analysis of Protein Targets
Check if the same lab product or an alternative is used in the 5 most similar protocols
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According to the manufacturer’s instructions, proteins were extracted using a total protein extraction kit (Keygen Biotech, Jiangsu) and detected with a bicinchoninic acid (BCA) protein assay kit (Keygen Biotech, Jiangsu). Proteins were resolved by SDS-PAGE and transferred to polyvinylidene difluoride membranes. These membranes were blocked with 5% fat-free milk in PBS-Tween 20 (Solarbio, Beijing) and incubated with primary antibodies against PTGER2 (1:1000; Abcam, USA), α-SMA (1:2000; Servicebio, Wuhan), Col-I (1:2000; Servicebio, Wuhan) or β-actin (1:3000; Servicebio, Wuhan). Membranes were then incubated with the proper horseradish peroxidase (HRP)-conjugated secondary antibodies (all at 1:3000; Servicebio, Wuhan). After subsequent washing, the immunoreactive bands were developed by enhanced chemiluminescence, exposed in the dark room and analyzed for densitometry.
3
Immunohistochemistry of Dorsal Skin
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Dorsal skin samples were embedded in OCT and sectioned at 8um thickness. Sections were fixed in formalin for 10mins, followed by two H2O washes (10mins each). Sections were blocked with blocking buffer (10% donkey serum in 1xPBST) for 1h. Primary antibodies were diluted in blocking buffer and applied to the tissue and incubated overnight at 4C. Secondary antibodies were diluted in 1xPBST for 1h at room temperature following three 1xPBST washes (5mins). Sections were washed in 1xPBST three times (5mins) and mounted with Fluoroshield with DAPI (Abcam, ab104139). Images were taken using the Leica DM7200 fluorescence imaging platform with LAS X version 3.7.5. Antibodies used: F4/80 1:600 (Biolegend, Cat#123101); Ly6G 1:600 (Biolegend, Cat#127601); CD3 1:800 (Biolegend, Cat#100201); Ccl2 1:100 (Novus, Cat#NBP1–42312); Ccl20 1:100 (Novus, Cat#AF760-SP); Cox-1 1:500 (Abcam, Cat#ab109025); Cox-2 1:600 (Cayman, Cat#160106); Ptges2 1:200 (Abclonal, Cat#A7137); Ptges3 1:200 (Abclonal, Cat#A5194); Ptger2 1:500 (Abcam, Cat#ab167171); Cd49f 1:100 (BD Biosciences, Cat#555734); Dct 1:600 (Abcam, Cat#ab221144), together with suitable Alex Fluor secondary antibodies (Abcam, Cat#ab150072, ab150149, ab150152, Fisher, Cat#A21207)
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