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
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1 MG
$951.00
About This Item
UNSPSC Code:
12352200
NACRES:
NA.77
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
solubility
soluble (Aqueous buffers or H2O. Please see vial label for lot-specific reconstitution volume.)
shipped in
ambient
storage temp.
2-8°C
Related Categories
1 of 4
This Item | 582231 | D0564 | 582243 |
|---|---|---|---|
| form solid | form lyophilized | form lyophilized powder | form lyophilized |
| manufacturer/tradename Calbiochem® | manufacturer/tradename Calbiochem® | manufacturer/tradename - | manufacturer/tradename Calbiochem® |
| Quality Level 100 | Quality Level 200 | Quality Level 200 | Quality Level 100 |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
| storage condition OK to freeze | storage condition OK to freeze | storage condition - | storage condition OK to freeze |
| shipped in ambient | shipped in ambient | shipped in - | shipped in - |
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
signalword
Danger
hcodes
Hazard Classifications
Acute Tox. 1 Inhalation - Acute Tox. 1 Oral
Storage Class
6.1A - Combustible acute toxic Cat. 1 and 2 / very toxic hazardous materials
wgk_germany
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’.
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how to reconstitute diphtheria toxin
1 answer-
Helpful?
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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?
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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.
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