To determine PEPT2-HA cell surface expression by chemiluminescence [53 (link)], defolliculated oocytes were first injected with 20 ng cRNA encoding either PEPT2-HA and/or 10 ng cRNA encoding USP18. After 4 days of incubation, oocytes were blocked with 1% BSA in ND96 solution for 20 minutes and then incubated with anti-HA-HRP antibody (diluted 1:1000, Miltenyi Biotec, Germany). Next, oocytes were washed three times 10 minutes each with 1% BSA in ND96 solution followed by three times 10 minutes each in ND96 solution. Individual oocytes were placed in 96 well plates with 20 μl of SuperSignal ELISA Femto Maximum Sensitivity Substrate (Pierce, Rockford, IL, USA), and chemiluminescence of single occytes was quantified in a luminometer (Walter Wallac 2 plate reader, Perkin Elmer, Juegesheim, Germany) by integrating the signal over a period of 1 s. Results display normalized relative light units.
Walter wallac 2 plate reader
Manufactured by PerkinElmer
Sourced in Germany
The Walter Wallac 2 plate reader is a multi-mode detection platform designed for various applications in life science research and drug discovery. It offers high-sensitivity detection of fluorescence, luminescence, and absorbance across 96- and 384-well microplates.
Automatically generated - may contain errors
3 protocols using walter wallac 2 plate reader
1
Quantifying PEPT2-HA Cell Surface Expression
2
EAAT2 Cell Surface Expression Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For detection of EAAT2 cell surface expression, defolliculated oocytes were incubated during 1hour at room temperature with rabbit anti-EAAT2 antibody (diluted 1:500, Alamone labs, Israel), washed and subsequently incubated with secondary, HRP-conjugated, goat anti-rabbit IgG antibody (1:1000, Cell Signaling technology, USA). After washing the second anti-body, oocytes were placed in 96 well plates for chemiluminescence assay with the SuperSignal ELISA Femto Maximum Sensitivity Substrate (Pierce, Rockford, IL, USA), following the manufacturer´s recommendations. Chemiluminescence of each oocyte was determined in a luminometer (Walter Wallac 2 plate reader, Perkin Elmer, Juegesheim, Germany) [36] .
3
Quantifying Oocyte Chemiluminescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cellular Physiology and Biochemistry with secondary, HRP-conjugated anti-mouse IgG antibody (1:2500, GE Healthcare Life Sciences). Individual oocytes were placed in 96 well plates with 20 µl of SuperSignal ELISA Femto Maximum Sensitivity Substrate (Pierce, Rockford, IL, USA) and chemiluminescence of single oocytes was quantified in a luminometer (Walter Wallac 2 plate reader, Perkin Elmer, Juegesheim, Germany) by integrating the signal over a period of 1 s. Results display normalized arbitrary light units. Integrity of the measured oocytes was assessed by visual control after the measurement to avoid unspecific light signals from the cytosol [73, 74] .
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!