The CEP tissues collected were fixed immediately with 4% formalin for 12 h and dehydrated for 48 h with 40% sucrose solution. Then, the CEP tissues were cut to 5 μm thickness and dried on glass slides for 3 h. A 3% H2O2 solution was used to stain the slides for 5 min. Then, the tissues were blocked with 3% serum and incubated with mouse anti-human O-GlcNAcylation antibody (RL2) (MA1-027, Thermo, MA, USA), mouse anti-human COL1 antibody (ab90395, Abcam, Cambridge, UK), and rabbit anti-human COL2 antibody (ab34712, Abcam), overnight at 4 °C. The tissue sections were washed three times in PBS and incubated in the corresponding secondary antibody (anti-rabbit immunoglobulin-horseradish peroxidase [IgG-HRP]-linked antibody, #7074, Cell Signaling Technology, MA, USA; anti-mouse IgG-HRP-linked antibody, #7076, Cell Signaling) for 1 h. Finally, nickel-diaminobenzidine images were obtained with a microscope.
Anti rabbit immunoglobulin horseradish peroxidase igg hrp linked antibody
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-rabbit immunoglobulin-horseradish peroxidase [IgG-HRP]-linked antibody is a secondary antibody that binds to rabbit primary antibodies. It is conjugated with horseradish peroxidase, an enzyme that can be detected using a colorimetric substrate.
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2 protocols using anti rabbit immunoglobulin horseradish peroxidase igg hrp linked antibody
1
Histochemical Analysis of CEP Tissue
2
Immunohistochemical Analysis of Hypoxia and Collagen
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The CEP specimens were freshly collected in the operating room, fixed immediately in 4% formalin for 12 hr, and dehydrated through 40% sucrose solutions for 48 hr. Sections were cut to 5-μm thickness, put onto glass slides, and dried for 3 hr. The slides for staining were blocked for endogenous peroxidase activity using 3% H2O2 solution for 5 min. They were then blocked with 3% goat serum, incubated in rabbit anti-human HIF1A antibody (Abcam, ab85886), mouse anti-human COL1 antibody (Abcam, ab90395), and rabbit anti-human COL2 antibody (Abcam, ab34712), respectively, overnight at 4°C, washed three times in PBS, and incubated in the corresponding secondary antibody (anti-rabbit immunoglobulin-horseradish peroxidase [IgG-HRP]-linked antibody, Cell Signaling Technology, #7074; anti-mouse IgG-HRP-linked antibody, Cell Signaling, #7076) for 1 hr with washing three times in PBS. Immunoreactive proteins were revealed with nickel-diaminobenzidine. Images were obtained with a microscope.
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