Forty-eight hours after the last training session and in an ad libitum state, 0.5 mL of venous blood were collected via cardiac puncture in all the rats. In these samples, plasma insulin was measured using an ELISA kit (Abcam, ab100578), and TNF-α determinations were carried out with the specific ELISA kit (Thermo SCIENTIFIC, ER3TNFA, Rockford IL.) according to the manufacturer’s instructions. Blood samples were stored in Eppendorf tubes with EDTA, centrifuged at 2000 rcf for 20 min, and the supernatant plasma was stored at -80°C for subsequent ELISA assays, according to the manufacturer’s instructions. Absorbance was measured at 450 nm (iMark microplate absorbance reader, BIO-RAD, Cat. 168-1130, Hercules, CA) 30 min after stop solution was added. The samples were measured in duplicate, and the mean absorbance was calculated. Detection limits were < 50 pg/mL for insulin and < 15 pg/mL for TNF-α. Inter and intra assay means coefficients of variance were < 10% and < 12% for insulin and < 10% and 10% for TNF-α.
Ab100578
Manufactured by Abcam
Sourced in United States, United Kingdom
Ab100578 is a lab equipment product manufactured by Abcam. It is designed to perform a specific function in a laboratory setting. The core function of this product is to [CORE FUNCTION DESCRIPTION]. No further details or interpretations about the intended use of this product are provided.
Automatically generated - may contain errors
Lab products found in correlation
Imark microplate absorbance reader, by Bio-Rad (1 mentions)
Er3tnfa, by Thermo Fisher Scientific (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Ab8304, by Abcam (1 mentions)
Ab150074, by Abcam (1 mentions)
Ab100768, by Abcam (1 mentions)
Insulin elisa kit, by Abcam (1 mentions)
Glucometer, by Roche (1 mentions)
Synergy ht reader, by Agilent Technologies (1 mentions)
Dgcg0, by R&D Systems (1 mentions)
Ab102523, by Abcam (1 mentions)
Ab46105, by Abcam (1 mentions)
Ab197742, by Abcam (1 mentions)
Ab222503, by Abcam (1 mentions)
9 protocols using ab100578
1
Plasma Insulin and TNF-α Analysis
2
Quantitative Proinsulin and Insulin Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Corresponding reagents were prepared and placed at room temperature. Sample or standard solution (100 µl) was added in the sample wells or standard wells, respectively. Corresponding antibodies, including proinsulin antibody (1:500; cat. no. ab8304; Abcam, Cambridge, MA, USA) and insulin antibody (1:500; cat. no. ab100578; Abcam) were then added for incubation at room temperature for a total of 2 h. Subsequently, horseradish peroxidase-labeled antibodies were added (1:1,000; cat. no. ab150074; Abcam). After incubation for 1 h at room temperature, 100 µl substrate was added, followed by colour reaction in the dark for 10 min. The absorbance at the wavelength of 450 nm was detected by a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).
3
Metabolic Effects of BD3526 in GK Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sixteen 18-week-old GK rats were used in this study. The rats were randomly divided into two groups of eight rats each. The rats in the BD3526 group received 2 ml of 50 mg/ml lyophilized BD3526 metabolites daily by gavage, whereas the rats in the control group received 2 ml of 50 mg/ml skim milk powder daily by gavage. The animals were individually caged with free access to a normal chewing bar and drinking water. The animals were kept alone in cages and allowed to eat freely. All animals were kept at 25°C with a 12-h light/dark cycle. The gavage experiments lasted for 4 weeks.
All rats were fasted for 8 h prior to administration of a glucose tolerance test. During the experiment, each rat was intraperitoneally injected with 5 g/kg of glucose solution. Blood glucose was monitored at 0, 15, 30, 60, and 120 min. Serum insulin was measured using an enzyme-linked immunosorbent assay (ELISA) kit (Abcam, ab100578) after the rats were killed. IL-1β was measured using an ELISA kit (Abcam, ab100768). The specific experimental methods were conducted in strict accordance with the manufacturer's instructions.
The experiments were performed in strict accordance with the experimental protocol. After dissection of the rats, the intestinal samples were immediately frozen at −80°C for long-term preservation prior to RNA extraction and cytokine detection.
All rats were fasted for 8 h prior to administration of a glucose tolerance test. During the experiment, each rat was intraperitoneally injected with 5 g/kg of glucose solution. Blood glucose was monitored at 0, 15, 30, 60, and 120 min. Serum insulin was measured using an enzyme-linked immunosorbent assay (ELISA) kit (Abcam, ab100578) after the rats were killed. IL-1β was measured using an ELISA kit (Abcam, ab100768). The specific experimental methods were conducted in strict accordance with the manufacturer's instructions.
The experiments were performed in strict accordance with the experimental protocol. After dissection of the rats, the intestinal samples were immediately frozen at −80°C for long-term preservation prior to RNA extraction and cytokine detection.
4
Rat Blood Glucose and Insulin Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were collected from the rat tail vein and the fasting blood glucose, as well as the blood insulin, were measured after 8 weeks using a glucometer (GlucoDr, South Korea) and enzyme-linked immunosorbent assay kit (insulin ELISA kit, Ab100578, Abcam, Cambridge, UK), respectively.
5
Quantifying Insulin Levels in Diabetic Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse and human insulin levels were quantified in plasma collected from mice with STZ-induced diabetes using ELISA kits specific for each species (mouse: 90,082, Crystal Chem, IL, USA; human: ab100578, Abcam, Cambridge, UK). Blood samples were collected from the tail vein in a non-fasting state at 15 days and 30 days using mini-vettes (50 μl, POCT, Nümbrecht, Germany). The plasma was obtained by incubating the blood for 1 h on ice, followed by centrifugation at 3000 g for 20 min; the separated plasma was then transferred to a new tube. The amounts of insulin in the plasma were quantified using the ELISA kit according to the manufacturer’s instructions.
6
Hormonal and Metabolic Profiling in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The morning after 2 weeks of treatment, blood samples were collected from the abdominal aorta of anesthetized rats using a procoagulation tube and centrifuged at 3,000 rpm for 20 min using the 4°C centrifuge (ICT15RE, Hitachi, Tokyo, Japan). Then the upper supernatants were carefully transferred to the EP tube using a pipette gun and immediately stored at a −80°C refrigerator for further detection. The levels of estradiol (E2, JEB-13691, Jinyibo, Wuxi, China), testosterone (T, ab10866, Abcam, Cambridge, MA, USA), LH (JEB-13706, Jinyibo, China), follicle-stimulating hormone (FSH; JEB-13680, Jinyibo, China), fasting insulin (FINS, ab100578, Abcam, USA), LPS (JEB-13984, Jinyibo, China), and tumor necrosis factor-α (TNF-α, ERC102a.96, Xinbosheng, China) were analyzed by ELISA kits, which were operated in strict accordance with each instruction manual. Blood glucose (Glu) was measured using Roche glucometer before anesthesia. The homeostasis model assessment of IR index (HOMA-IR) and HOMA of beta-cell function index (HOMA-beta) was calculated using the following formulas:
7
Plasma Enzyme and Insulin Measurements
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma concentrations of alanine aminotransferase (ALT) [S.G.P.T ERBA kit, 120903; Baddi, India] and aspartate aminotransferase (AST) [S.G.O.T ERBA kit, B081717; Baddi, India] were measured with an enzymatic color test according to the manufacturer’s protocols and were measured using an ERBA semi-automated biochemistry analyzer. Plasma insulin was measured using a rat enzyme-linked immunosorbent assay kit [ab100578, Abcam; Cambridge, MA, USA].
8
Comprehensive Metabolic Biomarker Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood glucose from the OGTT time points, creatinine, and a lipid panel were determined using standard colorimet-ric procedures (Biosystems, Spain). Anti-glutamic acid decarboxylase (anti-GAD65) antibodies were measured using a biotin-coupled sandwich ELISA (Kronus, ID, USA, CAT# KR7710), with an analytical sensitivity of 0.18 U/mL and functional sensitivity (the lowest level yielding an interassay coefficient of variation not >20%) of 4 U/mL. Absorbance was read at a wavelength of 450 nm according to the manufacturer’s instructions. We measured plasma insulin at times 0, 30, 60, and 120 of the OGTT using a sandwich ELISA (Abcam, CAT# ab100578) with a functional sensitivity of 4 µUI/mL. Absorbance for the insulin ELISA was also read at a wavelength of 450 nm according to the manufacturer’s instructions. HbA1c was measured by the boronate affinity method,11 (link) using the NycoCard II® reader. This technique and manufacturer have been certified by the United States National Glycohemoglobin Standardization Program as traceable to the Diabetes Control and Complications Trial standard. All ELISA plates were read in a BioTek Synergy HT Reader®. All measurements were performed in duplicate, and positive and negative controls were ran within each batch. We executed all procedures at the Diabetes, Lipids and Metabolism laboratory of Uni-versidad de los Andes.
9
Serum Cytokine and Hormone Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice serum was separated and stored at −80 °C for ELISA. The amylase levels in the serum were measured using the amylase assay colorimetric kit (ab102523) from Abcam (Cambridge, UK), and the level of amylase was recorded in mU/ml (nmol/min/ml). The serum glucagon levels were assessed using the glucagon ELISA kit (DGCG0) from R&D Systems (Minneapolis, MN, USA). Glucagon values are presented in pg/ml. Serum insulin levels were calculated using a commercial insulin ELISA kit (ab100578) from Abcam (Cambridge, UK). Insulin values are presented in μlU/ml. ELISA kits (ab197742, ab222503, and ab46105) for detection of the proinflammatory cytokines IL-1β, IL-6, and TNF-α were obtained from Abcam (Cambridge, UK) and used according to the manufacturer’s instructions, and the levels of IL-1β, IL-6, and TNF-α were recorded in pg/ml.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!