Key features and details
- Expression system: Native
- Purity: > 95% SDS-PAGE
- Active: Yes
- Suitable for: SDS-PAGE, Functional Studies
Product nameNative Clostridium difficile Toxin B protein
Measured by vercytotoxicity assay with 2 pg/ml as lowest point at which rounded cells are observed.
Purity> 95 % SDS-PAGE.
Clostridium difficile Toxin B has been highly purified.
Protein lengthFull length protein
Predicted molecular weight270 kDa
Amino acids2 to 2366
Our Abpromise guarantee covers the use of ab124001 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
This protein is derived from a pathogenic organism, and may be involved in a disease process, consequently exposure may have adverse health effects.
Functional Studies Protocol
1.1 Vero cells are plated out the night before on a 96-well plate at a concentration of 20K cells per well (in 100ml), then incubated at 37°C, 5% CO2. The following day they should be lightly confluent.
1.2 Lyophilised toxin/toxoid is resuspended as per the instructions in the certificate of analysis, and diluted in duplicate to 0.1mg/ml in DMEM (200ml total) without FBS in the first well of a non-TC-treated 96-well plate. A serial 5-fold dilution is performed (40ul in 160ul) in the same media using a multichannel for 11 dilutions in total, leaving the 12th dilution as a media control.
1.3 20ul of each dilution is then pipetted onto each well of the pre-prepared Vero cells and then incubated at 37°C, 5% CO2 for 24 hours.
1.4 The following day, the cells are examined using an inverted phase contrast tissue culture microscope. Cells with active toxin on should be rounded, cells without significant toxicity should look like the control cells.
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Store at -20°C or -80°C. Avoid freeze / thaw cycle. Please see notes section.
Constituents: 1.19% HEPES, 5% Sucrose, 0.88% Sodium chloride
This product is an active protein and may elicit a biological response in vivo, handle with caution.
ReconstitutionReconstitute in 250µl sterile distilled water. Reconstituted liquid can be stored at -80°C or -20°C to extend stability.
- C. difficile Toxin B
- Clostridium difficile Toxin B
- Cytotoxin B
RelevanceClostridium difficile Toxin B (tcdB or ToxB) is a glucosyltranferase which is known to innactivate Rho, Cdc42 and Rac within target cells. This toxin is encoded on a pathogenicity region of the C. difficile chromossome and is expressed during the log and stationary phases of growth in response to a variety of environmental stimuli.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab124001 has been referenced in 4 publications.
- Magnotti F et al. Pyrin dephosphorylation is sufficient to trigger inflammasome activation in familial Mediterranean fever patients. EMBO Mol Med 11:e10547 (2019). PubMed: 31589380
- Hartley-Tassell LE et al. Lectin activity of the TcdA and TcdB toxins of Clostridium difficile. Infect Immun N/A:N/A (2018). PubMed: 30530621
- Moghaddas F et al. A novel Pyrin-Associated Autoinflammation with Neutrophilic Dermatosis mutation further defines 14-3-3 binding of pyrin and distinction to Familial Mediterranean Fever. Ann Rheum Dis 76:2085-2094 (2017). PubMed: 28835462
- Hutton ML et al. Bovine antibodies targeting primary and recurrent Clostridium difficile disease are a potent antibiotic alternative. Sci Rep 7:3665 (2017). Neutralising ; Clostridium difficile . PubMed: 28623367