C. difficile Toxin B/TcdB Antibody

Detection ofC. difficileToxin B/TcdB by Western Blot.

Western blot shows recombinantC. difficileToxin B/TcdB (Catalog # 6246-GT). PVDF membrane was probed with 1 µg/mL of Sheep Anti-C. difficileToxin B/TcdB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6246) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Toxin B/TcdB at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of C. difficile Toxin B/TcdB by Western Blot

Effective TcdB cell interactions correlate with endocytosis. (A, B) Representative confocal maximum-intensity projections of CHO-K1 cells stained with 250 nM Alexa Fluor 647-labeled TcdB1 B2′B3 (red) (A) or Alexa Fluor 647-labeled TcdB2 B2′B3 (red) (B). Ten micromolar calcein AM (green) was used to counterstain the cytoplasm of live cells with intact membranes, and 0.5 μg/ml Hoechst 33258 (blue) was used to visualize the nucleus. (C) Quantification of the images in panels A and B, expressed as corrected total cell fluorescence. (D) Representative immunoblotting of full-length TcdB1 and TcdB2 associated with CHO-K1 cells in the presence or absence the dynamin inhibitor dynasore (80 μM). Incubations were carried out for 30 min at 37°C. (E) Quantification of data presented in panel D, expressed as relative band density. (F) Representative immunoblotting of full-length TcdB1 and TcdB2 associated with CHO-K1 cells when incubations were carried out for 30 min at a temperature that permits (37°C) or inhibits (ice) endocytosis. (G) Quantification of data presented in panel F, expressed as relative band density. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28776043), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of C. difficile Toxin B/TcdB by Western Blot

Mechanism-of-action of niclosamide inhibition of TcdB intoxication. c In vitro auto-processing assay. Compound or DMSO was added to 100 nM TcdB, and pre-incubated for 60 min. To initiate auto-processing, InsP6 was added (100 μM) and incubated @ 37 °C for 20 min before stopping with Laemlii sample buffer. Niclosamide at concentrations up to 10 μM did not affect auto-processing (n = 2). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30531960), licensed under a CC-BY license. Not internally tested by R&D Systems.