Serum measurements were carried out as follows: ferritin (Abcam Mouse Ferritin ELISA #ab157713, Abcam, Cambridge, MA, USA); insulin (CrystalChem Ultrasensitive Mouse Insulin ELISA #90080, Elk Grove Village, IL, USA); leptin (Quantikine Mouse/Rat Leptin ELISA #MOB00, Minneapolis, MN, USA); adiponectin (Abcam Mouse ELISA #ab108785, Cambridge, MA, USA); and a lipid panel with transaminases (Piccolo® Lipid Panel Plus, Greisham, Germany). Hepatic measurements with colorimetric kits include the following: triglyceride (Cayman Chemical #10010303, Ann Arbor, MI, USA); protein carbonyl content (Abcam #ab126287, Cambridge, MA, USA); and citrate synthase activity (Cayman Chemical #701040, Ann Arbor, MI, USA). For citrate synthase activity measurement in human biopsy livers (Sigma Aldrich #CS0720, Poole, UK), an activity assay kit was used.
Ab126287
Manufactured by Abcam
Sourced in United States
Ab126287 is a primary antibody product offered by Abcam. It is a purified monoclonal antibody that can be used for laboratory research applications. The core function of this product is to serve as a detection reagent for a specific target.
Automatically generated - may contain errors
Lab products found in correlation
Ab118970, by Abcam (2 mentions)
Cs0720, by Merck Group (1 mentions)
Ultra sensitive mouse insulin elisa, by Crystal Chem (1 mentions)
T5500, by Merck Group (1 mentions)
Synergy htx, by Agilent Technologies (1 mentions)
Mak187, by Merck Group (1 mentions)
Ab113851, by Abcam (1 mentions)
Ab201734, by Abcam (1 mentions)
Ab65329, by Abcam (1 mentions)
Mak085, by Merck Group (1 mentions)
Ab178020, by Abcam (1 mentions)
Protein carbonyl assay kit, by Abcam (1 mentions)
Glycogen assay kit, by Merck Group (1 mentions)
Ab27642, by Abcam (1 mentions)
Elisa kit, by Abcam (1 mentions)
Ab102536, by Abcam (1 mentions)
11 protocols using ab126287
1
Comprehensive Serum and Hepatic Biomarker Analysis
2
Biochemical Marker Quantification Protocol
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The separated serum was used to estimate some biochemical markers such as liver function tests, e.g., alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), gamma-glutamyl transferase (γGT), total protein, albumin, globulin, total bilirubin, and kidney function tests, e.g., creatinine, urea, and cystatin C. Besides, quantitative measurements of carbonyl protein content and malonaldehyde (MDA) were performed to evaluate protein and lipid peroxidation, respectively. All biochemical tests listed above were completed using a fully automated biochemical analyzers (HumaStar 600, Human Diagnostics Worldwide, Germany) except cystatin C and carbonyl protein contents that were quantified according to the instructions provided by the kit manufacturer (ab119590, Abcam, UK) and (ab126287, Abcam, UK), respectively, using a 96-well plate reader (Synergy HTX, BioTeK, USA). The evaluation of lipid peroxidation was performed by the determination of malondialdehyde (MDA) concentration in plasma samples, according to Lima et al. [17 (link)]. A serum sample of 250 μl was mixed with 500 μl of 0.6% 2-thiobarbaturic acid (TBA) (T5500, Sigma Aldrich, USA).
3
Protein Carbonylation Quantification
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Protein carbonylation was determined using a DNPH (2,4-dinitrophenylhydrazine)-based colorimetric assay kit (ab126287 - Abcam) following manufacturer's recommendations.
4
Antioxidant Capacity Evaluation in Brain
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5
Antioxidant Status in Cortical Brain Tissue
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The homogenate of cortical brain tissue was used to measure total antioxidant capacity, lipid peroxidation, and protein carbonylation. The concentrations of total antioxidant capacity (Sigma-Aldrich, MAK187), malondialdehyde (MDA, Sigma-Aldrich, MAK085), and protein carbonyl content (Abcam, ab126287) were measured according to the manufacturer’s instructions. The concentration of total antioxidants was calculated as nmol per μL, MDA was calculated as nmol per mg protein, and the protein carbonyl content was calculated as nmol per mg protein. All measurements were performed by a person who was blinded to the grouping.
6
Oxidative Stress and Lipid Peroxidation in S. aureus Induced by CBLEO
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Oxidative stress and lipid peroxidation of S. aureus induced by CBLEO were assessed by detecting the amounts of ROS generation and protein oxidation (two key markers of oxidative stress) and MDA accumulation (one biomarker of lipid peroxidation) [7 (link),71 (link)]. The S. aureus cells (107 CFU/mL) were incubated with different doses (1/2×MIC, 1×MIC, and 2×MIC) of CBLEO at 37 °C, and the samples were collected respectively at different times (0−8 h). After centrifuging at 10,000 rpm for 10 min under 4 °C, the obtained cell pellets were used to determine protein carbonyl, ROS, and MDA by using assay kits of ab126287, ab113851, and ab118970 (Abcam, Shanghai, China), respectively. The amount of ROS was given in fold of untreated controls, and the content of protein carbonyl (protein oxidation product) was standardized to the used amount of S. aureus cells and defined as nmol/mg. The MDA content was standardized to the used amounts of S. aureus cells, and the result was expressed as nmol/mg.
7
Antioxidant and Oxidative Stress Markers
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Human plasma was isolated from total blood collected from the first day of recruitment to the last day of exercise training. Total antioxidant capacity, 8-hydroxy-2′-deoxyguanosine (8-OHdG), and carbonyl levels were measured using commercial kits (ab65329, ab201734, and ab126287; Abcam) according to the manufacturer’s instructions. The malondialdehyde (MDA) level was determined by a commercial kit (MAK085, Sigma) according to the manufacturer’s instructions.
8
Measurement of Protein Carbonylation
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Protein carbonyl content in the lung tissue lysates or in mitochondrial proteins were measured photometrically following derivatization with DNPH using a commercial kit (ab126287) from Abcam (Cambridge, MA, USA). In a similar set of experiments carbonylated proteins were also detected following immunoblotting using an anti-DNPH antibody (Abcam, ab178020).
9
Cardiac Glycogen and Protein Carbonyl
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Cardiac glycogen content was measured using a glycogen assay kit according to manufacturer’s protocol (Sigma). Units were calculated using molecular weight of 180.16 g/mol. Protein Carbonyl content was assessed using a protein carbonyl assay kit according to manufacturer’s protocol and ∼45 mg tissue was utilized (abcam; ab126287).
10
Immunological and Oxidative Biomarkers in Rabbits
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ELISA kits provided by Elabscince Company (Houston, TX, 77079, USA) were used to assess the levels of immunoglobulins (IgA and IgM) in the serum of rabbits [32] following the directions protocols. The serum levels of malondialdehyde (MDA; ab27642) were assessed based on the reaction with thiobarbituric acid [33] and protein carbonyl (PC; ab126287) were assessed using commercial ELISA kits (Abcam company) based on the reaction with DNPH [34] . Glutathione (GSH) and superoxide dismutase (SOD) were assessed using commercial kits provided by Nanjing Jiancheng Bioengineering Institute (Nanjing, China), following the recommendations in the pamphlet. The serum level of interferon-gamma (IFN-γ; ab273238) was assessed using a colorimetric technique with a commercial diagnostic kit (Abcam company), which had a sensitivity of 0.16 ng/mL [35] . The levels of serum triiodothyronine (T3) and corticosterone were measured using commercially available assays. T3 levels were determined using a direct ELISA assay from Diametra Srl, Segrate, Italy. Corticosterone levels were determined using corticosterone ELISA kits [36] , following the manufacturer's recommendations (Assay Pro-LLC, Saint Charles, MO, USA).
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