Anti AKT, phospho Thr308  

Immunohistochemistry of Mouse anti-AKT pT308 antibody. Tissue: human brain cerebellum tissue (40X). Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pT308antibody at 20 μg/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: staining of Purkinje neurons and cell processes in the cerebellum, cytosolic as well as occasionally nuclear. Staining: AKT pT308 as precipitated red signal with hematoxylin purple nuclear counterstain.

Western Blot of Mouse anti-AKT pT308 antibody. Lane 1: non-phosphorylated AKT in untreated NIH/3T3 cells (p/n W10-000-358). Lane 2: phosphorylated AKT on PDGF stimulated NIH/3T3 cell lysates (p/n W10-001-377). Load: 15 μg per lane. Primary antibody: AKT pT308 antibody at a 1:4,000 dilution in TBS with 3% BSA, for 3 h at 4° C. Secondary antibody: Peroxidase conjugated Gt-a-Mouse IgG (Fc) (p/n 610-1303) was used at a 1:40,000 dilution for 1 h at 4° C. Block: 3% BSA (p/n BSA-30) in TBS for 30 min at RT. Predicted/Observed size: (indicated by arrowhead at ~56 kDa). Other band(s): unspecific.

PHB and phospho-PHB (pT258) colocalize with AKT and phospho-AKT in the mitochondrial sheath of mouse mature sperm. Colocalization of (a) PHB (magenta) or (b) phospho-PHB (pT258; magenta) with AKT (green) or phospho-AKT (pT308, pS473; green), and the mitochondrial sheath (red) of mouse cauda epididymal sperm, was determined using a mitochondrial membrane potential probe (MITO) and confocal fluorescence microscopy. The inserts showed the image amplified for “Line Scan” colocalization analysis. The plot profiles of MITO (red), PHB (magenta) or phospho-PHB (pT258; magenta), AKT (green) or phospho-AKT (pT308, pS473; green) colocalization along the region of interest (ROI) were constructed and analyzed using LAS AF Lite software, as shown in the last column on the right. The negative controls (the last rows of a and b) were run with rabbit (R-IgG) and mouse (M-IgG) IgG isotypes, instead of specific primary antibodies. Scale bars = 20 μm. For amplified image (insert), scale bars = 5 μm. PHB: prohibitin; AKT: protein kinase B; DAPI: 4',6-diamidino-2-phenylindole. Fig 1. PMID: 32859869