LDH Cytotoxicity Assay Kit (Colorimetric) (CAT#: CB-P150-K) Datasheet

Product Type
Kit
Description
The LDH assay kit (cytotoxicity) uses WST to quickly and sensitively detect LDH released from damaged cells.

The LDH assay (also known as the LDH release assay) is a cell death/cytotoxicity assay used to assess the level of plasma membrane damage in a cell population. Lactate dehydrogenase (LDH) is a stable enzyme that exists in all cell types. Once the plasma membrane is damaged, it will be quickly released into the cell culture medium. LDH is the most widely used marker for performing cytotoxicity assays.

The LDH analysis procedure is based on an enzymatic coupling reaction: LDH released from cells oxidizes lactic acid to produce NADH, and then NADH reacts with WST to produce yellow. The intensity of the color produced is directly related to the number of lysed cells.

This assay requires only 10 µl of culture medium, so the background of serum and culture medium is significantly reduced. The cells can be cultured in a medium containing 10% normal serum. The assay does not require reducing serum or special media.

In addition, since WST is very stable, the reaction can be read multiple times, and the reaction can be stopped at any point during the reaction.

LDH activity can be easily quantified at OD450nm by spectrophotometer or microplate reader.
Applications
Functional Studies
Size
1 kit
Detection Method
Colorimetric/Fluorometric
Sample
Cell culture media
Species Reactivity
Mammals
Protocol Length
1h 00m
Components
ell Lysis Solution 1 x 5ml
LDH (Positive control) 1 vial
LDH Assay Buffer 1 x 50ml
Stop Solution 1 x 5ml
WST Substrate Mix 1 vial
Requires
Microplate reader
Handling Advice
LDH assay protocol summary:
- transfer 10µl culture medium into new plate
- add LDH reaction mix and incubate for 30 min at room temp
- analyze with microplate reader
Storage
Please refer to protocols.
Shelf Life
12 months
Research Use
For research use only. Not for human or drug use

Figure 1 LDH Cytotoxicity Assay


Figure 2 LDH Cytotoxicity Assay
Jurkat T cells were cultured in 96-well plate in 100 μl of culture medium. LDH Assay was performed using 10μl of culture medium using the WST probe. Low control (white bar); High control (black bar).


Figure 3 LDH Cytotoxicity Assay
Comparison of WST-1 and INT based LDH assays. 3T3 cells were cultured in a 96-well plate in 100 µl of culture medium. The LDH assay was performed using 10 µl of culture medium using WST-1 (Brown bar) and INT (Green bar) methods. The WST-1 based LDH assay is more stable and sensitive than the INT based method.

All products and services are for Research Use Only. Do Not use in humans.

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