Monoclonal anti hif 1α h1alpha67

HIF-1α was immunostained on 5-μm paraffin-embedded tissue sections using the streptavidin-peroxidase technique. Briefly, paraffin sections were deparaffinized, hydrated, and subjected to antigen retrieval and blocking of endogenous peroxidase activity. Slides were incubated with the primary antibody monoclonal anti-HIF-1α (H1alpha67; 1:200; Novus Biologicals, Littleton, CO) overnight at 4 °C, followed by incubation with non-biotinkit (EliVision™ superKIT-9922, Maixin, Fuzhou, China) reagents A and B for 15 min each to allow the horseradish peroxidase polymer to bind to the primary antibody. Diaminobenzidine was used for staining, followed by a hematoxylin counterstain.
The expression of HIF-1α in the renal tubular epithelial cells was observed by optical microscopy with a ×400 magnification visual field and blinded scored as 0–4 points: 0 points, weakly expressed or without nuclear staining; 1 point, positive rate of nuclear staining <25%; 2 points, positive rate of nuclear staining of 25–50%; 3 points, positive rate of nuclear staining of 50–75%; and 4 points, positive rate of nuclear staining >75%.