Human SHARPIN Antibody

Detection of Human SHARPIN by Western Blot.

Western blot shows lysates of U2OS human osteosarcoma cell line and THP-1 human acute monocytic leukemia cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human SHARPIN Monoclonal Antibody (Catalog # MAB8100) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for SHARPIN at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of SHARPIN by Western Blot

SHARPIN affects the induction of IFN response. (A) PLC3WT and PLC3SHARPIN-KO cells (indicated, respectively, with + and −), are mock- or HEV ORF3-transfected (indicated, respectively, with − and +). Successful knockout and ORF3 transfection verified by Western blot. B-actin is displayed as a control. Two replicates of each condition are displayed. (B) PLC3WT and PLC3SHARPIN-KO cells were mock- or HEV ORF3-transfected and two days later transfected with poly (I:C). IFN-beta (left) and IFN-lambda mRNA levels measured by qRT-PCR. Transcripts normalized to reference genes (ATP5B, CYC1, YWHAZ, HPRT1, RPL30) and scaled to PLC3WT—MOCK (ORF3-). (C) PLC3WT and PLC3SHARPIN-KO cells infected with HEV (MOI 22000) and infection levels analyzed 6 days later by HEV ORF2 immunostaining. Representative confocal images on the right. Nuclei in blue and ORF2 in green. Results are represented as mean ± SEM compiled from three independent experiments. Statistical significance calculated by (multiple) t-test. Differences were considered as statistically significant when p < 0.05 (*). Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1999-4915/15/12/2412), licensed under a CC-BY license. Not internally tested by R&D Systems.