The largest database of trusted experimental protocols

Ridascreen verotoxin microtiter plates

Manufactured by R-Biopharm
Sourced in Germany

The RIDASCREEN Verotoxin microtiter plates are a laboratory test kit designed to detect the presence of verotoxins (also known as Shiga-like toxins) in biological samples. The plates contain pre-coated wells that can be used to perform enzyme-linked immunosorbent assays (ELISA) to identify verotoxin-producing bacteria, such as Escherichia coli (E. coli). The core function of the product is to provide a standardized and reliable method for the detection of verotoxins, which are important markers for the identification of pathogenic bacteria.

Automatically generated - may contain errors

2 protocols using ridascreen verotoxin microtiter plates

1

Quantitative Sandwich-ELISA for Stx2

Check if the same lab product or an alternative is used in the 5 most similar protocols
Sandwich-ELISA was used to detect Stx2 in the bacterial culture supernatants, and was carried out in RIDASCREEN Verotoxin microtiter plates (R-Biopharm GmbH, Darmstadt, Germany) coated with capture antibodies for detecting both Stx1 and Stx2. A monoclonal antibody against Stx2 (LSBio, WA, USA) conjugated with horseradish peroxidase using Peroxidase Labeling Kit–NH2 (Dojindo, Kumamoto, Japan) was used as the detection antibody. A standard curve was determined with 2-fold serial dilutions of purified Stx2 (provided by Denka Seiken Co. Ltd.). Detection was carried out with urea peroxidase/TMB substrate reagent and 1 M sulfuric acid as the stop reagent, both were provided in the RIDASCREEN Verotoxin kit. Absorbance at 450 nm (A450) was measured using a 96-well plate reader (Multiscan FC, Thermo Fisher Scientific, MA, USA).
+ Open protocol
+ Expand
2

Quantification of Shiga Toxins in Bacterial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
We inoculated bacterial cells into 40 mL of CAYE broth (Merck, Darmstadt, Germany) and grew them to mid-log phase at 37°C with shaking. We then added mitomycin C (MMC) to the culture at a final concentration of 500 ng/mL. After MMC addition, we collected 100 μL of the culture every hour and immediately subjected it to sonication using a Bioruptor (Cosmo Bio, Tokyo, Japan). We obtained the soluble fractions of each cell lysate via centrifugation at 14,000 × g for 10 min at 4°C. We determined Stx1 and Stx2 concentrations in each cell lysate using a previously described sandwich ELISA (40 (link)). We captured Stx using RIDASCREEN Verotoxin microtiter plates (R-Biopharm, Darmstadt, Germany) coated with capture antibodies that recognize both Stx1 and Stx2. We conjugated monoclonal antibodies against Stx1 and Stx2 (LSBio, Seattle, WA, USA) with horseradish peroxidase using the Peroxidase Labeling Kit–NH2 (Dojindo, Kumamoto, Japan) and employed them as detection antibodies. We used reagents supplied in the RIDASCREEN Verotoxin kit for detection. Finally, we measured the absorbance at 450 nm (A450) using Tecan Infinite 200 PRO (Tecan, Männedorf, Switzerland).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!