Mice were euthanized by carbon dioxide asphyxiation. Tumors were dissected and fixed with 10% formalin overnight, transferred to 70% ethanol, and then embedded in paraffin. 3μM thick sections were cut and stained with H&E. Immunohistochemistry on paraffin-embeded sections was performed using antibodies against Nqo1 (1:100, Sigma, HPA007308), phospho-histone-H3 (1:100, Ser10; Cell Signaling, #9701), 8-oxo-deoxyguanosine (1:100, Abcam, #ab48508), and cleaved caspase3 (1:200, Cell Signalin, #9579). Chromogenic IHC was performed on a Leica Bond RX and stained slides were imaged on a Leica SCN400 F whole-slide scanner. For Nqo1, 8-oxo-deoxyguanosine, and phospho-histone-H3 staining antigen retrieval was performed using antigen retrieval buffer pH6 (Leica) for 20 min and for cleaved caspase3 staining, antigen retrieval buffer pH9 (Leica) was used for 20min. For detection, Leica Bond Polymer Refine Detection secondary antibody (Leica, #DS9800) was used according to manufacturer’s protocol for Nqo1, phosphor-histone-H3 and cleaved caspase3. 8-oxo-deoxyguanosine was detected with the Biocare MM HRP-polymer secondary antibody (Biocare, #MM620H) according to manufacturer’s protocol.
Bond polymer refine detection secondary antibody
Manufactured by Leica
The Leica Bond Polymer Refine Detection secondary antibody is a laboratory reagent used in immunohistochemistry (IHC) applications. It is designed to enhance the detection and visualization of primary antibodies bound to target antigens in tissue samples.
Automatically generated - may contain errors
3 protocols using bond polymer refine detection secondary antibody
1
Immunohistochemical Characterization of Tumor Samples
2
Lung Tumor Histological Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were euthanized by carbon dioxide asphyxiation. Lung with tumors were dissected, infused, and fixed with 10% formalin overnight; transferred to 70% ethanol; and then embedded in paraffin. Sections (3 mM thick) were cut and stained with H&E. Immunohistochemistry on paraffin-embedded sections was performed using antibodies against Nqo1 (1:100; Sigma-Aldrich, HPA007308) and G6pd (1:5000; Abcam, ab993). Chromogenic IHC was performed on a Leica Bond RX, and stained slides were imaged on a Leica SCN400 F whole-slide scanner. For Nqo1 and G6pd staining, antigen retrieval was performed using antigen retrieval buffer pH 6 (Leica) for 20 min. For detection, Leica Bond Polymer Refine Detection secondary antibody (Leica, no. DS9800) was used according to the manufacturer’s protocol for Nqo1 and G6pd.
3
Quantitative analysis of NQO1 in lung tumors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were euthanized by lethal doses of ketamine and xylazine. Lungs were inflated and incubated overnight at room temperature (RT) with 10% formalin. They were subsequently transferred to 70% ethanol, and subsequently embedded in paraffin. 5uM sections were stained with H&E or subjected to other immunohistochemical staining. For immunohistochemistry (IHC) we used antibody against NQO1 (1:100, HPA007308, Sigma-Aldrich). Immunohistochemistry was performed on a Leica Bond RX and slides were imaged on a Leica SCN400F whole slide scanner. For NQO1, antigen retrieval was performed using antigen retrieval buffer pH=6 (Leica) for 20min. For detection, Leica Bond Polymer Refine Detection secondary antibody was used according to manufacturer’s protocol. Total tumor lung area was quantified via H&E-stained slides using QuPath software.103 (link) Tumor burden and IHC analyses were done in a blinded fashion.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!