Anti qsox1 antibody

Lentiviruses packaged with short hairpin RNA (shRNA) constructs specific for human QSOX1 (sh528, sh742) or a nonspecific scrambled sequence (shScr) were produced in 293T cells as reported by Katchman et al. [2 (link)]. Knockdown of QSOX1 in MIAPaCa-2 was confirmed by western blotting using anti-QSOX1 antibody (Protein Tech, Chicago, IL USA).

Briefly, after antigen retrieval, tissue sections were incubated with a rabbit polyclonal anti-QSOX1 antibody (1:200 dilution; Proteintech, Chicago, IL, USA) overnight at 4°C, followed by incubation with a biotinylated secondary antibody and an avidin-biotin peroxidase complex (ZSGB-Bio, Beijing, China). Then, the immune reactions were developed by adding DAB chromogen- substrate solution (ZSGB-Bio, Beijing, China) to the slides. Harris hematoxylin was used for counterstaining. Negative controls were run in parallel with all reactions. All specimens were scored by a board-certified pathologist. The scoring was determined as follows: i) the proportion of tumor cells with IHC staining for QSOX1 protein expression (0: no staining, 1 (Low): <33%, 2 (Intermediate): 33 to 66%, 3 (High): >66%), and ii) the intensity of the stain (0: negative, 1: weak, 2: moderate, 3: strong staining intensity). An overall staining score of ≤3 indicated low expression and a score of >3 was considered high expression.