Immunohistochemistry for detection of the LC3 protein and the macrophage marker F4/80 was carried out using standard protocols. Briefly, unstained sections of lungs or tumors were deparaffinized and rehydrated, and then incubated in the peroxidase blocking reagent (BioVision cat #K405‐50). Antigen retrieval was performed by boiling the sections in sodium citrate for 20 min. To decrease background staining, the slides were incubated for 1 h in the mouse on mouse blocking reagent (Vector Labs MKB‐2213‐1), followed by overnight incubation with the primary antibodies (LC3B Cell Signaling 3868S rabbit polyclonal, 1:300; or F4/80 Cell Signaling 70076S rabbit monoclonal, 1:200). Next day, the slides were washed and incubated with One‐Step HRP polymer (BioVision cat #K405‐50) for 30 min at RT. The slides were then washed several times and then incubated with the DAB chromogen for 10 min at RT, followed by several washing steps and quick counterstain with Hematoxylin. Slides were then mounted and visualized under the upright microscope (Nikon).
One step hrp polymer
Manufactured by Abcam
The One-Step HRP Polymer is a horseradish peroxidase (HRP) conjugate used for immunohistochemistry and immunocytochemistry applications. It is designed to provide reliable and consistent signal amplification during the detection process.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using one step hrp polymer
1
Immunohistochemical Detection of LC3 and F4/80 in Lung and Tumor Tissues
2
Immunohistochemical Analysis of Autophagy and Macrophages
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Immunohistochemistry for detection of the LC3 protein and the macrophage marker F4/80 was carried out using standard protocols. Briefly, unstained sections of lungs or tumors were deparaffinized and rehydrated, and then incubated in the peroxidase blocking reagent (BioVision cat #K405-50). Antigen retrieval was perfomed by boiling the sections in sodium citrate for 20 min. To decrease background staining, the slides were incubated for 1 h in the mouse on mouse blocking reagent (Vector Labs MKB-2213-1), followed by overnight incubation with the primary antibodies (LC3B Cell Signaling 3868S rabbit polyclonal, 1:300; or F4/80 Cell Signaling 70076S rabbit monoclonal, 1:200). Next day, the slides were washed and incubated with One-Step HRP polymer (BioVision cat #K405-50) for 30 min at RT. The slides were then washed several times and then incubated with the DAB chromogen for 10 min at RT, followed by several washing steps and quick counterstain with Hematoxylin. Slides were then mounted and visualized under the upright microscope (Nikon).
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