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Rabbit Recombinant Monoclonal Clostridium difficile Toxin A antibody. Carrier free. Suitable for sELISA and reacts with Clostridium difficile samples.

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Images

Sandwich ELISA - Anti-Toxin A antibody [EPR23359-20] - BSA and Azide free (Detector) (AB316226), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
sELISA
Clostridium difficile
Tested

Tested
Tested

Species

Clostridium difficile

Dilution info

-

Notes

-

Target data

Function

Toxin APrecursor of a cytotoxin that targets and disrupts the colonic epithelium, inducing the host inflammatory and innate immune responses and resulting in diarrhea and pseudomembranous colitis (PubMed:20844489). TcdA and TcdB constitute the main toxins that mediate the pathology of C.difficile infection, an opportunistic pathogen that colonizes the colon when the normal gut microbiome is disrupted (PubMed:19252482, PubMed:20844489). Compared to TcdB, TcdA is less virulent and less important for inducing the host inflammatory and innate immune responses (PubMed:19252482). This form constitutes the precursor of the toxin: it enters into host cells and mediates autoprocessing to release the active toxin (Glucosyltransferase TcdA) into the host cytosol (By similarity). Targets colonic epithelia by binding to some receptor, and enters host cells via clathrin-mediated endocytosis (By similarity). Binding to LDLR, as well as carbohydrates and sulfated glycosaminoglycans on host cell surface contribute to entry into cells (PubMed:16622409, PubMed:1670930, PubMed:31160825). In contrast to TcdB, Frizzled receptors FZD1, FZD2 and FZD7 do not act as host receptors in the colonic epithelium for TcdA (PubMed:27680706). Once entered into host cells, acidification in the endosome promotes the membrane insertion of the translocation region and formation of a pore, leading to translocation of the GT44 and peptidase C80 domains across the endosomal membrane (By similarity). This activates the peptidase C80 domain and autocatalytic processing, releasing the N-terminal part (Glucosyltransferase TcdA), which constitutes the active part of the toxin, in the cytosol (PubMed:17334356, PubMed:19553670, PubMed:27571750).Glucosyltransferase TcdAActive form of the toxin, which is released into the host cytosol following autoprocessing and inactivates small GTPases (PubMed:22267739, PubMed:22747490, PubMed:24905543, PubMed:30622517, PubMed:7775453). Acts by mediating monoglucosylation of small GTPases of the Rho family (Rac1, RhoA, RhoB, RhoC, Rap2A and Cdc42) in host cells at the conserved threonine residue located in the switch I region ('Thr-37/35'), using UDP-alpha-D-glucose as the sugar donor (PubMed:22267739, PubMed:22747490, PubMed:24905543, PubMed:30622517, PubMed:7775453). Monoglucosylation of host small GTPases completely prevents the recognition of the downstream effector, blocking the GTPases in their inactive form, leading to actin cytoskeleton disruption and cell death, resulting in the loss of colonic epithelial barrier function (PubMed:7775453). Also able to catalyze monoglucosylation of some members of the Ras family (H-Ras/HRAS, K-Ras/KRAS and N-Ras/NRAS), but with much less efficiency than with Rho proteins, suggesting that it does not act on Ras proteins in vivo (PubMed:30622517).

Alternative names

toxA, tcdA, Toxin A

Recommended products

Rabbit Recombinant Monoclonal Clostridium difficile Toxin A antibody. Carrier free. Suitable for sELISA and reacts with Clostridium difficile samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR23359-20

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Notes

ab316226 is a BSA and Azide free antibody supplied in an unconjugated
format and it is suitable for sandwich ELISAs to quantify Clostridioides difficile Toxin A.
The recommended pair for sandwich ELISA is:

Capture: Anti-Toxin A antibody [EPR23359-22] - BSA and Azide free (Capture) ab316225, Clostridioides difficile Toxin A Capture Antibody (unconjugated)

Detector: ab316226, Clostridioides difficile Toxin A Detector Antibody (unconjugated)

The reference range value is 78-5000 pg/mL.

The recommended antibody orientation is based on internal optimization
for ELISAbased assays. Antibody orientation is assay dependent and
needs to be optimized for each assay type. Please note that the
range provided for this antibody is only an estimation based on the
performance of the product using the recommended antibody pair.
Performance of the antibody pair will depend on the specific
characteristics of your assay. We guarantee the product works in
sandwich ELISA, but we do not guarantee the sensitivity or dynamic
range of the antibody in your assay.

Our carrier-free formats are supplied in a buffer free of BSA,
sodium azide and glycerol for higher conjugation efficiency.
The antibodies are provided at an approximate concentration of
1 mg/ml as measured by the protein A280 method. Use our conjugation
kits for antibody conjugates that are ready-to-use in as
little as 20 minutes with <1 minute hands-on-time and 100% antibody
recovery: available for fluorescent dyes, HRP, biotin and gold.

The recommended antibody orientation is based on internal optimization for ELISA-based assays. Antibody orientation is assay dependent and needs to be optimized for each assay type. Please note that the range provided for this antibody is only an estimation based on the performance of the product using the recommended antibody pair. Performance of the antibody pair will depend on the specific characteristics of your assay. We guarantee the product works in sandwich ELISA, but we do not guarantee the sensitivity or dynamic range of the antibody in your assay.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Clostridium difficile Toxin A also known as C. diff toxin A or enterotoxin A acts mechanically by disrupting the cytoskeleton of intestinal epithelial cells. Weighing approximately 308 kDa this toxin gets expressed by Clostridium difficile bacteria during infection. C. diff toxin A enters the host cells inactivating Rho family GTPases leading to cell rounding and apoptosis. This disruption contributes to the weakening of the gut lining increasing its permeability.

Biological function summary

The main role of C. diff toxin A involves causing inflammation in the colonic mucosa. It functions as part of a dual toxin system with Toxin B. Toxin A binds to receptors on the surface of intestinal cells initiating a cascade that recruits neutrophils and other immune cells. This immune response exacerbates tissue damage and results in the observed symptoms of C. diff infections. The interaction with Toxin B enhances its pathogenic capacity further altering the tight junctions within the gut epithelium.

Pathways

C. diff toxin A affects the Rho-GTPase signaling pathway by acting as a glucosyltransferase and modifying small GTPases like Rho Rac and Cdc42. These changes impair the cell's cytoskeletal dynamics and disrupt cellular functions related to structural maintenance. Furthermore C. diff toxin A also influences the NF-kB pathway which plays a role in inflammation and immune responses. Toxin A's role in modifying these pathways highlights the coordinated interplay with Toxin B promoting inflammation and cytotoxicity.

Associated diseases and disorders

The role of C. diff toxin A is prominent in Clostridium difficile infections (CDI) a significant cause of antibiotic-associated diarrhea and colitis. The presence of C. diff toxin A connects strongly to the severity of CDI leading to prolonged illness and complications like pseudomembranous colitis. Studies indicate an interaction with proteins like Toxin B that enhances the severity of infection and tissue destruction. Understanding C. diff toxin A's mechanisms can assist in developing therapeutic strategies targeting both toxins for effective treatment of CDI.

Product promise

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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