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TOX7 Sigma

In Vitro Toxicology Assay Kit, Lactic Dehydrogenase based

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Biochem/physiol Actions

LDH reduces NAD+, which then converts a tetrazolium dye to a soluble, colored formazan derivative.

Application

Tox7 is a member of Sigma’s CytoToxicity Kit series. It is based on the measurement of lactate dehydrogenase (LDH) release from cells as a measure of cell membrane damage and cytotoxicity. This technique has been utilized as an alternative to 51Cr release for cell mediated cytotoxicity assays, as well as conventional cytotoxicity resulting from interaction of a test material with the cell.

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Safety & Documentation

Safety Information

Symbol 
GHS05  GHS05
Signal word 
Danger
Hazard statements 
Precautionary statements 
RIDADR 
NONH for all modes of transport

Frequently Asked Questions

Can TOX7, In Vitro Toxicology Assay kit be used to quantitate the amount of LDH in my samples?
This assay kit was designed as a semi-quantitative assay for LDH. The change in the OD between the treated samples and control samples is to be compared and is generally reported as a fold increase/decrease from the control. The kit does not contain a LDH standard. Product L3916 can be used as an LDH standard for this assay. The detection range (LDH units/ml) for the kit has not been established.
I'm using the TOX7 In Vitro Toxicology Assay Kit.  I do not have a 690 nm filter to use for a reference/background.  What can I use?
The reference wavelength needs to be greater than the absorbance spectra of the assay. The greater the difference between the assay wavelength and the reference wavelength, the better.  A reference wavelength of 590 nm or greater can be used with the substrate for this assay.  If a reference wavelength is not available, the assay will work.  The use of the reference wavelength increases the signal to noise ratio to give a better assay range.
The TOX7 dye solution has a precipitate present. Can I still use it?
This information can be found on TOX7 Product Information Sheet.Product No. L2277 LDH Assay Dye Solution is a saturated solution and some precipitation may appear upon thawing. The product will partially re-solubilize when warmed to 37°C and the resulting solution can be decanted away from any remaining precipitate. Precipitated dye added to reactions, or removed from the solution will not affect the performance or results obtained from this kit.  Click on the following link to view TOX7 Product Information Sheet:  http://www.sigmaaldrich.com/sigma/bulletin/TOX7bul.pdf
I am using a protease inhibitor cocktail in the cell culture media.  Will this interfere with the TOX7 In Vitro Toxicology assay?
The protease inhibitor cocktails do not interfere with LDH activity.  Media from cells cultured with protease inhibitors can be used in this assay.
Will Triton X100  interfere with the TOX7 In Vitro Toxicology Kit assay?
Triton X100 will not interfere with TOX7 assay.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
Can you use tissue explants in product TOX7, In Vitro Toxicology Assay Kit, Lactic Dehydrogenase based?
Unfortunately, we have not tested product TOX7, In Vitro Toxicology Assay Kit, on tissue samples.  If the tissue is minced well and can be lysed completely with the lysis buffer for the cells, it may work to determine cell biomass.  The following paper modified the assay for use with explants. Benyo, D.F., et al., "Hypoxia Stimulates Cytokine Production by Villous Explants from the Human Placenta," The Journal of Clinical Endocrinology & Metabolism 82(5), 1582-1588 (1997). Assessment of explant viability was routinely monitored by measuring the release of LDH into medium relative to a 1% Triton X-100 (Sigma)-lysed positive control. The assay employed a modification of the TOX7 viability kit supplied by Sigma-Aldrich and entailed adding a 20-fold volume excess of assay mixture (160 mL) to the medium sample tested (8 mL). The assay mixture consisted of equal volumes of substrate (lactate), enzyme (diaphorase), and dye (nitro blue tetrazolium) solutions, and the reaction was allowed to proceed for 20 min in a microtiter plate before the addition of 0.1 vol 1N HCl. Color change was quantified by reading the absorbance at 490 nm, and the percent cell death calculated.
What is the minimum number of cells to use in the LDH assay, Product TOX7, In Vitro Toxicology Assay Kit?
Using Product TOX7, In Vitro Toxicology Assay Kit, the minimum cell number for best response is 2 x 104.
  What kind of solution is the A5955 - Antibiotic Antimycotic Solution (100×), Stabilized product suspended in?
A5955 - Antibiotic Antimycotic Solution (100×), Stabilized is supsended in a sodium citrate buffer solution. Source: Israel production group
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Protocols & Articles

Articles

Cell Viability and Proliferation Assays

Assays to measure cellular proliferation, cell viability, and cytotoxicity are commonly used to monitor the response and health of cells in culture after treatment with various stimuli. The proper ch...
Article Based on
BioFiles v6 n5, 17–21
Keywords: Cancer, Cell proliferation, Click chemistry, Enzyme-linked immunosorbent assay, Flow cytometry, Glycolysis, Immunocytochemistry, Immunohistochemistry, Microscopy, Nucleic acid denaturation, Reductions, Solvents, Western blot

Peer-Reviewed Papers
15

References

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