Animal studies were performed in accordance with Washington University IACUC. Mice were randomly designated for STZ treatment and transplantation groups. Mouse number per group was selected to allow for statistical significance based on our prior studies (15 (link), 17 (link), 18 (link)). Surgical procedures and follow up studies were performed by unblinded individuals. Male 7 wk old NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice were purchased from Jackson Laboratories, rendered diabetic with injection of 45 mg/kg STZ (R&D) for 5 d, with diabetes confirmed after 8 d. Anaesthetized mice were injected with 5x106 WS4unedit stage 6 cells, 5x106 WS4corr stage 6 cells, 5x106 (4000 IEQ) islet cells, or saline under the kidney capsule. Islet transplantation was performed in a separate cohort. Animals were monitored up to 6 months. Blood glucose was measured with a Contour Blood Glucose Monitoring System (Bayer). Glucose tolerance and in vivo GSIS assays were performed by fasting mice for 4 hr and injecting with 2 g/kg glucose. Serum hormones were quantified using Human Ultrasensitive Insulin ELISA kit (ALPCO Diagnostics), Mouse C-peptide ELISA (ALPCO Diagnostics), and human proinsulin ELISA (Mercodia). 12-wk after transplantation, live nephrectomy was performed on 2 anaesthetized transplanted mice.
Human ultrasensitive insulin elisa kit
Manufactured by ALPCO
Sourced in United States
The Human Ultrasensitive Insulin ELISA kit is a diagnostic tool used to measure insulin levels in human samples. It is a quantitative enzyme-linked immunosorbent assay (ELISA) designed for the detection and quantification of insulin in serum, plasma, and other biological fluids.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using human ultrasensitive insulin elisa kit
1
Evaluating Stem Cell Transplant Efficacy
2
SC β Cell Insulin Secretion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SC β cell differentiation between 28 and 35 days was sampled in discs (1.15 × 105 cells/mm2). Cell discs and SC β clusters were washed with PBS followed by preincubation in low (2.8 mM) glucose Krebs buffer for 2 hours to remove residual insulin. Discs and SC β clusters were then washed with PBS twice and incubated in low-glucose Krebs buffer for 30 min, and then the supernatant was collected. The discs and clusters were further washed with PBS twice and incubated in high glucose (20 mM) Krebs buffer for 30 min, and the supernatant was collected. These sequences were repeated for two additional times. Last, discs and clusters were incubated with 30 mM KCl (depolarization challenge) in PBS for 30 min, and then the supernatant was collected. These clusters were next dispersed into single cells using TrypLE Express (Gibco) for cell counting. Supernatant samples containing insulin were processed using the Human Ultrasensitive Insulin ELISA Kit (ALPCO Diagnostics, Salem, NH, USA) and normalized based on live cell numbers.
3
Glucose-Stimulated Insulin Secretion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analysis was conducted following previously described procedure (48 (link)). The ceMEDs were loaded 150 µL of MIN6 cells or SC-βCs at a seeding density with 3 million cells or 2,000 IEQ. For SC-βCs, the cells were extracted from rat subcutaneous space 14-d posttransplantation and then measured using GSIS analysis. After overnight culture, the devices were removed from culture media and starved in 1.5-mL 2.8 mM glucose (Millipore Sigma) solution for 2 h. The devices were then sequentially submerged in solutions of 2.8 mM glucose, 20 mM glucose, 2.8 mM glucose, and 30 mM KCl for either 30 or 60 min each. Small aliquots of the solutions (10 µL) were collected at each time point and insulin content was quantified by mouse insulin enzyme-linked immunosorbent assay (ELISA) (Mercodia) and human ultrasensitive insulin ELISA kit (ALPCO Diagnostics). Then, the cells were collected and dispersed with TryplE (15 min), stained them with Trypan blue, and the glucose content was normalized to the number of live cells.
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!