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Merck
CN

TOX7

Sigma-Aldrich

体外毒性检测试剂盒,基于乳酸脱氢酶

别名:

LDH cytotoxicity assay

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About This Item

UNSPSC代码:
12352207
NACRES:
NA.84

形式

liquid

质量水平

用途

 kit sufficient for 500 tests

包装

pkg of 1 kit

储存条件

dry at room temperature

技术

protein quantification: suitable

λmax

490 nm

应用

cell analysis
detection

检测方法

colorimetric

储存温度

−20°C

一般描述

Tox7是Sigma细胞毒性试剂盒系列产品的一部分。它通过对细胞释放的乳酸脱氢酶(LDH)的测定,从而对细胞膜损伤和细胞毒性进行测定。该技术已作为51Cr释放的替代方法,用于细胞介导的细胞毒性,以及由于测试物质与细胞相互作用而产生的常规细胞毒性测定中。

应用

基于乳酸脱氢酶的体外毒理学检测试剂盒已用于:
  • 检测人脐静脉内皮细胞(HUVEC)和人心肌细胞(HCM)中乳酸脱氢酶的释放,从而进行乳酸脱氢酶(LDH)测定
  • 确定MDA-MB-231细胞乳酸脱氢酶泄漏的活性
  • 检测LDH在RAW 264.7细胞 中的活性,以及用于在人类细胞系的LDH细胞毒性测定中
  • 用于分光光度法活细胞测定。在490 nm波长下检测转化染料的吸光度。

生化/生理作用

乳酸脱氢酶测定是一种通过细胞质总乳酸脱氢酶(LDH)或细胞膜完整性(以释放到培养基中的胞质LDH量表征)来测量细胞数量的方法。该方法简单、准确,并且结果可重复。该测定是基于LDH对NAD的还原。所得还原的NAD(NADH)用于将四唑鎓染料化学计量转化为可溶有色的甲瓒衍生物。
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危险声明

预防措施声明

危险分类

Aquatic Chronic 3

储存分类代码

10 - Combustible liquids

法规信息

常规特殊物品

分析证书(COA)

输入产品批号来搜索 分析证书(COA) 。批号可以在产品标签上"批“ (Lot或Batch)字后找到。

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  1. 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.

  2. 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.

  3. The TOX7 dye solution has a precipitate present. Can I still use it?

    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.

  4. 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.

  5. Will Triton X100  interfere with the TOX7 In Vitro Toxicology Kit assay?

    Triton X100 will not interfere with TOX7 assay.

  6. 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.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. 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.

  9. 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. 

  10. 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.

  11. 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.

  12. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

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