This material is soluble in aqueous buffers or water. The recommended resonstitution volume is reported on the product label. Please see the link below to review addtional information available in the product datasheet:
https://www.emdmillipore.com/product/Diphtheria-Toxin-Unnicked-Corynebacterium-diphtheriae-Calbiochem,EMD_BIO-322326#anchor_PDS
322326
Diphtheria Toxin, Unnicked
from Corynebacterium diphtheriae, Major band under reduced conditions (SDS-PAGE), <15% nicking, solid, protein synthesis inhibitor, Calbiochem®
Synonym(s):
DTx
Sign Into View Organizational & Contract Pricing
All Photos(1)
About This Item
UNSPSC Code:
12352200
NACRES:
NA.77
Pricing and availability is not currently available.
Recommended Products
Product Name
Diphtheria Toxin, Unnicked, Corynebacterium diphtheriae, Diphtheria toxin catalyzes ADP-ribosylation of eukaryotic aminoacyltransferase II (EF2) using NAD as substrate. Activation requires nicking with a protease followed by reduction with DTT.
Quality Level
description
RTECS - XW5807200
form
solid
manufacturer/tradename
Calbiochem®
storage condition
OK to freeze
shipped in
ambient
storage temp.
2-8°C
General description
Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae catalyzes ADP-ribosylation of eukaryotic aminoacyltransferase II (EF2) using NAD as substrate, thereby inhibiting protein synthesis. May also induce inter-nucleosomal breakdown. Causes DNA fragmentation and cytolysis in U937 cells. Activation requires nicking with a protease followed by reduction with DTT. Diphtheria toxin, synthesized and excreted as a proenzyme, is composed of a single polypeptide chain of 63 kDa. For its enzymatic activity to be expressed, the toxin must undergo two covalent alterations in structure. First, a mild proteolysis results in the formation of an enzymatically inactive "nicked toxin," which consists of two major fragments (A and B) linked by a disulfide bond. Reduction of the nicked toxin with thiols (DTT) releases the enzymatically active N-terminal A fragment (24 kDa). The C-terminal B fragment (39 kDa) has no apparent enzymatic activity. The B fragment is required for toxicity and is responsible for recognizing and binding of the toxin to cell surface receptors.
Application
Diphtheria Toxin, Unnicked, Corynebacterium diphtheriae has been used for intraperitoneal administration in mice for cell ablation.[1]
Biochem/physiol Actions
Cell permeable: no
Note: Toxicity may vary by lot of toxin. Each laboratory should determine the optimum dosage for each particular application.
Primary Target
Eukaryotic aminoacyltransferase II (EF2)
Eukaryotic aminoacyltransferase II (EF2)
Product does not compete with ATP.
Reversible: no
Warning
Toxicity: Harmful (C)
Physical form
Lyophilized from sterile 10 mM Tris, 1 mM EDTA, pH 7.5.
Reconstitution
Following reconstitution, aliquot, quickly freeze on dry ice, and freeze (-70°C). Subsequent thawing should be carried out only at room temperature. For assays employing very low concentrations of diphtheria toxin, the use of a carrier protein, such as BSA or HSA, is recommended.
Please see vial label for lot-specific reconstitution volume.
Analysis Note
Major band (under reducing conditions) of ~63 kDa
Other Notes
Diphtheria toxin is purified from Corynebacterium diphtheriae Park Williams strain 8 by a modified method of Pappenheimer, et al. As assessed by disc electrophoresis run at alkaline pH under non-denaturing conditions, this preparation migrates as a major band at 63 kDa, corresponding to the intact toxin. Two faster, more lightly stained bands (24 and 39 kDa), corresponding to A and B fragments, may be observed. Following trypsin treatment in DTT, diphtheria toxin exhibits high activity when assayed for its ability to ADP-ribosylate EF2. The ED50 for CHO cells is determined to be about 0.4 ng/ml.
Diphtheria toxin is purified from Corynebacterium diphtheriae Park Williams strain 8 by a modified method of Pappenheimer, et al. As assessed by disc electrophoresis run at alkaline pH under non-denaturing conditions, this preparation migrates as a major band at 63 kDa, corresponding to the intact toxin. Two faster, more lightly stained bands (24 and 39 kDa), corresponding to A and B fragments, may be observed. Following trypsin treatment in DTT, diphtheria toxin exhibits high activity when assayed for its ability to ADP-ribosylate EF2. The ED50 for CHO cells is determined to be about 0.4 ng/ml.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Acute Tox. 1 Inhalation - Acute Tox. 1 Oral
Storage Class Code
6.1A - Combustible acute toxic Cat. 1 and 2 / very toxic hazardous materials
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
Tamar Licht et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 40(5), 974-995 (2020-01-22)
Multiple insults to the brain lead to neuronal cell death, thus raising the question to what extent can lost neurons be replenished by adult neurogenesis. Here we focused on the hippocampus and especially the dentate gyrus (DG), a vulnerable brain
José Almeida-Santos et al.
European journal of immunology, 50(3), 439-444 (2019-11-16)
It is well established that therapeutic impairment of Foxp3+ Treg in mice and humans favors immune rejection of solid tumors. Less explored is the impact Foxp3 allelic variants may have on tumor incidence, progression and therapy. In this work, we
Weiqiang Jing et al.
International immunopharmacology, 78, 106012-106012 (2019-12-23)
Macrophages are recognized as one of the major cell types in tumor microenvironment, and macrophage infiltration has been predominantly associated with poor prognosis among patients with breast cancer. Using the murine models of triple-negative breast cancer in CD169-DTR mice, we
Keren Bahar Halpern et al.
Nature communications, 11(1), 1936-1936 (2020-04-24)
The intestinal epithelium is a structured organ composed of crypts harboring Lgr5+ stem cells, and villi harboring differentiated cells. Spatial transcriptomics have demonstrated profound zonation of epithelial gene expression along the villus axis, but the mechanisms shaping this spatial variability
-
how to reconstitute diphtheria toxin
1 answer-
Helpful?
-
-
For in vivo application on mice, is nicking necessary when using this product?
1 answer-
The Diphtheria Toxin preparation is not tested for in vivo application. However, based on data published in the literature, the unnicked form of the toxin seems to work as well as the nicked form for in vivo application.
Helpful?
-
-
Could information be provided on the suitability of product 322326-1MG for cell deletion involving DTT receptors in animals? Additionally, what are the differences between 322326-1MG and D0564?
1 answer-
Product 322336 has not been validated for in vivo application, but it has been known to work well on animal models with DT receptors. For further reference, find the citation "Curr Biol. 2021 Jun 7;31(11):2469-2476. Efferocytosis by Paneth cells within the intestine" with PMID: 33852873. 322326-1MG and D0564 products are same but with slight differences in specifications tested for batch release
Helpful?
-
Active Filters
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service
