Duodenum sections from six rats in each experimental group were deparaffinized, rehydrated, boiled in 10 mM citrate buffer (pH 6.0) for antigen retrieval, and endogenous peroxidase was quenched with 3% hydrogen peroxide. Subsequently, sections were incubated overnight at 4 °C with the following primary antibodies: anti‐proliferating cell nuclear antigen (PCNA), anti‐NPC1L1, anti‐ACAT2, anti‐MTP, and anti‐LXR (Santa Cruz Biotechnology, Quimigen, Madrid, Spain), and anti‐ABCG5, anti‐ABCG8 (Biorbyt Ltd., Quimigen, Madrid, Spain). After several washes, sections were incubated with biotinylated secondary antibody followed by streptavidin–biotin‐conjugated horseradish peroxidase (Sigma Aldrich, Madrid, Spain), using 3,3′‐diaminobenzidine (DAB) (Sigma Aldrich, Madrid, Spain) as substrate and then counterstained with Harris’ hematoxylin. Protein expression was evaluated by their staining pattern: weak 1), moderate 2), diffuse 3), or intense 4). The PCNA labeling index (PCNA‐LI) was calculated according to the following formula:
Streptavidin biotin conjugated horseradish peroxidase
Manufactured by Merck Group
Sourced in Spain
Streptavidin–biotin‐conjugated horseradish peroxidase is a molecular complex composed of streptavidin, a protein derived from the bacterium Streptomyces avidinii, conjugated to biotin-labeled horseradish peroxidase, an enzyme commonly used in various immunoassays and biochemical applications. This conjugate exhibits a high binding affinity between streptavidin and biotin, a property that is often utilized in research and diagnostic procedures.
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Lab products found in correlation
2 protocols using streptavidin biotin conjugated horseradish peroxidase
1
Immunohistochemical Analysis of Intestinal Lipid Metabolism
2
Quantifying Hepatic Receptor Expression
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Paraffin-embedded liver sections from five rats in each experimental group were deparaffinized and rehydrated in a graded ethanol series. Then, they were heated in 10 mM citrate buffer ( pH 6.0) for antigen retrieval, and endogenous peroxidase was quenched with 3% hydrogen peroxide. Subsequently, the sections were incubated overnight at 4 °C with the mouse primary antibodies anti-SR-B1 and anti-LDLr, anti-InsRbeta, anti-PI3K, and anti-pAKT Ser473 (Santa Cruz Biotechnology Quimigen, Madrid, Spain). After several washes, the sections were incubated with biotinylated secondary antibody followed by streptavidin-biotin-conjugated horseradish peroxidase (Sigma Aldrich, Madrid, Spain) using 3,3′-diaminobenzidine (DAB) (Sigma Aldrich, Madrid, Spain) as a substrate. The sections were then counterstained with Harris' haematoxylin. A total of 10 fields per section per rat (20× magnification for image analysis) were selected and analysed. The positive staining intensity was calculated by assigning scores to the stained area relative to the total field assessed.
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