Proliferative beta-cells were identified by insulin-Ki67 double immunofluorescence staining with rabbit anti-Ki67 (1:200; Thermo Fisher), guinea pig anti-insulin antibody (1:100; Abcam) and relevant secondary antibodies (1:1000; Abcam). Apoptotic beta-cells were identified by insulin and terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) (TMR red, Roche, Mississauga, ON, Canada) dual labeling. Results are expressed as the percentage of Ki67+, or Tunel+ beta-cells.
Relevant secondary antibodies
Manufactured by Abcam
Sourced in Canada, United Kingdom
Relevant secondary antibodies are laboratory reagents used to detect and visualize primary antibodies in various applications such as Western blotting, immunohistochemistry, and flow cytometry. They are designed to bind to the constant regions of primary antibodies, effectively amplifying the signal and enabling the detection of target proteins or molecules.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti ki67, by Thermo Fisher Scientific (2 mentions)
Tmr red, by Roche (2 mentions)
Guinea pig anti insulin antibody, by Abcam (1 mentions)
Rabbit anti pdx 1, by Cell Signaling Technology (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
3 protocols using relevant secondary antibodies
1
Proliferative and Apoptotic Beta-Cells
2
Quantifying Pancreatic Beta Cell Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proliferative beta cells were detected in the pancreatic sections by double immunofluorescence staining with guinea pig anti-insulin (1:1000; DAKO), rabbit anti-Ki67 (1:200; Thermo Fisher, Burlington, ON, Canada) antibodies, and relevant secondary antibodies (1:1000; Abcam). Regenerative beta cells were detected by double immunofluorescence staining with guinea pig anti-insulin (1:1000; DAKO), rabbit anti-PDX-1 (1:400; Cell Signaling Technology, Danvers, MA, United States) antibodies, and relevant secondary antibodies (1:200; Jackson ImmunoResearch Laboratories, West Grove, PA, United States). Apoptotic beta cells were also identified in pancreatic sections with insulin and terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) labeling (TMR red, Roche, Mississauga, ON, Canada) (Wang and Brubaker, 2002 (link); Robertson et al., 2008 (link)). Results are expressed as the percentage of Ki67+, PDX-1+, or Tunel+ beta cells. All immunofluorescent images were captured by an Olympus upright BX50 fluorescence microscope (Olympus, Richmond Hill, ON, Canada) at ×40 magnification.
3
Western Blot Protein Analysis in U87 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The assay was performed as previously described (18 (link)). Briefly, total protein was extracted from the U87 cells and concentration was measured using the bicinchoninic acid protein assay kit (Thermo Fisher Scientific, Inc., Waltham, MA, USA) according to the manufacturer's instructions. Protein samples (100 µg) were run on an SDS-polyacrylamide gel (3 h at 70 V) and the proteins were then transferred to nitrocellulose membranes (Sigma-Aldrich). The membranes were blocked with 5% bovine serum albumin (Sigma-Aldrich) for 2 h at room temperature and incubated with primary antibodies overnight at 4°C. This was followed by incubation with relevant secondary antibodies (1:10,000; Abcam, Cambridge, UK) for 1 h at room temperature. The bands were visualized using a ChemiDoc MP Imaging System (Bio-Rad Laboratories, Inc. (model 1708280; Hercules, CA, USA).
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!