All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: LACKW8-100 (for 100T)

Cat. No.: LACKW8-500 (for 500T)

Description

LDH Cytotoxicity Assay Kit with WST-8, also known as the LDH Assay Kit with WST-8 or LDH Release Assay Kit with WST-8, is a pre-packaged assay kit utilized for detecting lactate dehydrogenase (LDH) activity released during cytotoxicity via a colorimetric reaction based on WST-8.

This assay kit can be used for routine LDH activity detection and is more commonly employed for cytotoxicity testing based on LDH release. Additionally, based on the overall LDH activity in cells, this kit can also be used for cell proliferation assays.

LDH is an enzyme present in most mammalian cells and is widely distributed in various tissues, with relatively high concentrations in the heart, skeletal muscles, and kidneys. It serves as an important indicator in clinical enzyme profiles and can aid in the diagnosis of myocardial diseases. LDH catalyzes the conversion of lactate to pyruvate, along with the conversion of NAD+ to NADH. As it is typically released during tissue damage, often due to loss of cell membrane integrity, LDH is considered a biomarker for cell necrosis, common injuries, and related diseases.

The destruction of cell membrane structures caused by processes like necrosis, apoptosis, or secondary necrosis may result in the release of enzymes, including LDH, into the extracellular environment. This release occurs both in cultured cells, where LDH is released into the culture medium, and in vivo, where LDH may enter the tissue microenvironment and possibly the bloodstream. By detecting the activity of LDH released from ruptured cell membranes, quantitative analysis of cytotoxicity can be achieved. LDH release is considered a critical indicator of cell membrane integrity and is widely used in cytotoxicity testing, serving as a safe and effective alternative to earlier methods involving the radioactive labeling of cells with 51Cr and subsequent measurement of 51Cr release to assess membrane integrity.

Various colorimetric methods are available for measuring LDH activity, such as the DNP method, INT method, and WST-8 method. The WST-8 method, employed in this assay kit, typically offers higher sensitivity, greater absorbance changes, and more accurate results compared to the INT method.

This assay kit can detect both D-LDH and L-LDH or their mixture. The basic principle of the assay involves the reduction of NAD+ to NADH by LDH, followed by the reaction of NADH with WST-8 to generate water-soluble formazan dye, which produces an absorbance peak at 450nm. The absorbance is linearly correlated with LDH activity.

Features

• This assay kit offers a more sensitive reaction and greater convenience in usage. The reagents in this kit are non-toxic to cells, so there is no need to remove the cell culture medium when using the kit. The detection working solution can be directly added to the cell culture wells, making it a one-step process. Alternatively, the assay can be performed without affecting the cells by testing the cell culture medium, known as the non-destructive method, allowing the cells to be used for other experiments.
• This assay kit can detect the activity of LDH in samples such as cell culture medium and cell lysates.

Storage

Store at -20ºC, valid for one year. The chromogenic solution should be protected from light. After thawing, the reagents can be stored short-term at 4ºC and are valid for 2-3 days.

Precautions

• Freezing may cause partial inactivation of lactate dehydrogenase (LDH) in the samples. Samples can be stored at 4ºC for 2-3 days. It is recommended to complete the assay on the same day after sample preparation.
• When detecting LDH in cell culture medium, serum contains LDH, which can increase background readings. Set up control wells without cells but with the same volume of cell culture medium to eliminate background noise during testing. The higher the serum content, the higher the background values. If feasible for the experiment, consider using inactivated serum to significantly reduce LDH activity in the serum and thus lower the background. If feasible for the experiment, use serum-free medium or medium with lower serum concentrations to effectively reduce the background LDH activity in the serum.
• Cell overgrowth, high cell density, excessive centrifugation speed, and large temperature differences inside and outside the incubator can all increase the release of LDH from cells. Additionally, different cell types may have varying levels of LDH content.
• Without the addition of the stop solution, absorbance values will gradually increase over time. After adding the stop solution, the color development can be stable for up to 48 hours.
• If absolute quantification of LDH activity is desired, users need to prepare their own LDH standard.
• This product is intended for scientific research use by professionals only and should not be used for clinical diagnosis or treatment, food, or pharmaceutical purposes. It should not be stored in ordinary residences.
• For your safety and health, please wear laboratory coats and disposable gloves during operation.

Only for research and not intended for treatment of humans or animals

Journals Using SBS Genetech Products                                       Universities Using SBS Genetech Products

SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory