Rat elisa kit

To confirm PEDF overexpression in PD-MSCs, human PEDF in cell culture supernatant was analyzed according to the instructions using the human PEDF ELISA kit (Abcam). IL-6 and IL-10 levels in rat serum (n = 6/group) were analyzed by rat ELISA kit (Abcam) to determine proinflammatory and anti-inflammatory factor levels. Samples and standards were added to appropriate wells. Then, each antibody was incubated at room temperature. These were aspirated and each well was washed four times. TMB solution was incubated and stop solution was added. The OD at 450 nm was measured by an Epoch microplate reader (BioTek, Winooski, VT, USA). Each concentration was assessed by the trend line equation. All reactions were performed in triplicate.

To determine the oxidized glutathione (GSSH), the frozen rat livers (~30 mg) were homogenized (1:10 w:v ratio) in the extraction buffer (supplied with the kit) and determined calorimetrically (Abbkine, Inc, Wuhan, China, Cat#: KTB1610). For the lipid peroxidation marker 4-Hydroxy-2-Nonenal (4-HNE), the frozen livers (~30 mg) were homogenized (1:20 w/v ratio) using a RIPA buffer containing SDS and determined using a rat ELISA kit (Abcam, Cambridge, UK, Cat#ab238538). For the classic liver fibrosis marker type-I collagen (COL-1), the frozen livers (~30 mg) were homogenized (1:9 w/v ratio) using PBS (0.01 M, pH = 7.4) and measured using specific rat ELISA kits (Novus Biologicals, Centennial, CO, USA, Cat# NBP2-75823). All procedures were performed according to the manufacturer’s instructions.

Levels of serum insulin was determined by Insulin Rat ELISA Kit (Invitrogen, ERINS). Levels of cholesterol and triglycerides, in serum and liver, were estimated by Cholesterol Assay Kit (Abcam ab65390) and Triglyceride Assay Kit (Zen-Bio STG-1-NC). Liver function enzymes, Alanine transaminase (ALT) and aspartate aminotransferase (AST), were measured by Rat ELISA Kit (Abcam ab234579 and MyBioSource MBS264975; respectively). Levels of urea and creatinine were determined by Rat ELISA Kit (MyBioSource MBS2600001 and MBS007289; respectively). The antioxidant capacity of prepared Mel-C/L nanoparticles, in vivo, was evaluated from measuring levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity in pancreatic tissue by Rat ELISA Kit (MyBioSource MBS268427, MBS266897 and MBS727547; respectively). Levels of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-12 were determined by Rat ELISA Kit (MyBioSource MBS843359, Biolegend 438206, and MyBioSource MBS2507061, respectively).