Human/Mouse/Rat PNUTS Antibody

Detection of Mouse and Rat PNUTS by Western Blot.

Western blot shows lysates of TS1 mouse helper T cell line and Y3-Ag rat myeloid cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat PNUTS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF21581) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for PNUTS at approximately 114 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 5.

Detection of Human PNUTS by Simple Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. Specific bands were detected for PNUTS at approximately 100-150 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat PNUTS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF21581). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of PNUTS by Western Blot

PNUTS is an aging-regulated protein and regulates senescence in endothelial cells. A HUVECs were cultured for 3 or 17 passages (P) and PNUTS expression was measured by qRT-PCR (n = 4). b Aortic intima was isolated from young (8–12 weeks old) and aged (18–20 months old) mice and immediately lysed for RNA isolation. Intima RNA was sequenced. c–f HUVECs were transfected with siRNAs targeting PNUTS or a control sequence. c PNUTS protein levels were determined by WB at 48 h after transfection, relative to alpha-tubulin. Densitometric quantification is depicted on the right (n = 7). d Changes in p21 expression were assessed by qRT-PCR. Expression values are relative to siControl-treated HUVECs and normalized to RPSA mRNA (n = 6). e The percentage of senescent HUVECs was analyzed by staining for senescence-associated beta-Galactosidase (SA-( beta-Gal) 72 h after transfection. Images were taken (4 fields per sample) and the percentage of senescent cells (blue) over the total number of cells in each field was calculated in n = 3 independent experiments, 2–3 biological replicates per group and experiment). f Cell proliferation was assayed by testing the incorporation of EdU after transfection (n = 4). *p < 0.05, **p < 0.01, ***p < 0.001. Error bars depict the standard error of the mean (SEM). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38714838), licensed under a CC-BY license. Not internally tested by R&D Systems.