Toxin B/TcdB Antibody

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.

• Reactivity: Ba-Cd
• Applications: WB
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Toxin B/TcdB Antibody Summary

• Immunogen: E. coli-derived recombinant C. difficile Toxin B/TcdB
  Ser2-Leu543
  Accession # P18177
• Specificity: Detects C. difficile Toxin B/TcdB in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: tcdB
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL

Reactivity Notes

C. difficile

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Toxin B/TcdB Antibody

• TcdB
• toxB

Background

Clostridium difficile is the leading cause of hospital-acquired diarrhea, known as C. difficile-associated disease. The estimated number of cases of C. difficile-associated disease exceeds 250,000 per year (1), with health care costs approaching US $1 billion annually (2). The major virulence factors produced by C. difficile are two toxins, TcdA and TcdB. Both toxins can monoglucosylate and inactivate Rho family small GTPases within target cells, leading to disruption of vital signaling pathways in the cell, subsequently causing diarrhea, inflammation, and damage of colonic mucosa (3, 4, 5). Both toxins have a similar tripartite structure comprised of an N‑terminal glucosyltransferase domain, a C-terminal receptor binding domain, and a small hydrophobic span possibly involved in toxin translocation (6). Our recombinant TcdB consists of the enzymatic domain. Both TcdA and TcdB also have potassium-dependent UDP-Glc hydrolase activity, which is essentially glucosyltransferase activity with water as the acceptor molecule (7). Under same conditions, UDP-glucose hydrolysis by TcdB occurs at a rate about 5-fold greater than that of TcdA.

1. Wilkins, T.D. and Lyerly, D.M. (2003) J. Clin. Microbiol 41:53.
2. Kyne, L. et al. (2002) Clin. Infect. Dis. 34:346.
3. Voth, D.E. and Ballard, J.D. (2005) Clin. Microbiol. Rev. 18:247.
4. Chaves-Olarte, E. et al. (1996) J. Biol. Chem. 271:6925.
5. Just I, et al. (1995) J. Biol. Chem. 270:13932.
6. Hammond, G.A. and Johnson, J.L. (1995) Microb. Pathog. 19:203.
7. Ciesla, W.P. Jr. and Bobak, D.A. (1998) J. Biol. Chem. 273:16021.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Toxin B/TcdB Antibody (AF6246)(6)

Publications using AF6246

• Siddiqi, U;Lunnemann, HM;Childress, KO;Shupe, JA;Rutherford, SA;Farrow, MA;Washington, MK;Coffey, RJ;Lacy, DB;Markham, NO; Inhibition of EGFR/ErbB does not protect against C. difficile toxin B bioRxiv : the preprint server for biology 2024-05-14 [PMID: 38798529] (Immunocytochemistry, Human)
• Ahmed UKB, Shadid TM, Larabee JL, Ballard JD. Combined and Distinct Roles of Agr Proteins in Clostridioides difficile 630 Sporulation, Motility, and Toxin Production mBio 2021-01-14 [PMID: 33443122]
• J Tam, S Icho, E Utama, KE Orrell, RF Gómez-Biag, CM Theriot, HK Kroh, SA Rutherford, DB Lacy, RA Melnyk Intestinal bile acids directly modulate the structure and function of C. difficile TcdB toxin Proc. Natl. Acad. Sci. U.S.A., 2020-03-09;0(0):. 2020-03-09 [PMID: 32152097] (Western Blot, Human)
• J Tam, T Hamza, B Ma, K Chen, GL Beilhartz, J Ravel, H Feng, RA Melnyk Host-targeted niclosamide inhibits C. difficile virulence and prevents disease in mice without disrupting the gut microbiota Nat Commun, 2018-12-07;9(1):5233. 2018-12-07 [PMID: 30531960] (Western Blot, Human)
• JJ Hunt, JL Larabee, JD Ballard Amino Acid Differences in the 1753-to-1851 Region of TcdB Influence Variations in TcdB1 and TcdB2 Cell Entry mSphere, 2017-08-02;2(4):. 2017-08-02 [PMID: 28776043] (Western Blot, Chinese Hamster)
• JL Larabee, SJ Bland, JJ Hunt, JD Ballard Intrinsic Toxin-Derived Peptides Destabilize and Inactivate Clostridium difficile TcdB MBio, 2017-05-16;8(3):. 2017-05-16 [PMID: 28512094] (Western Blot, Hamster)

Bioinformatics

• Gene Symbol: tcdB
• Uniprot:
  • P18177(C. difficile)