LDH-Glo™ Cytotoxicity Assay

Monitor Cytotoxicity from Small Numbers of Cells

The LDH-Glo™ Cytotoxicity Assay is a bioluminescent plate-based assay for quantifying lactate dehydrogenase (LDH) release into the culture medium upon plasma membrane damage. The bioluminescent detection is more sensitive than colorimetric or fluorescent methods, allowing accurate detection of LDH from a small number of cells, including primary cells and 3D cell cultures.

This LDH assay involves removing only a small amount of cell media (2–5µl) from each treated well, allowing you to get more data by sampling the same well over time, and by using the remaining media and cells for other cell-based assays.

How the Assay Works

LDH released from damaged cells catalyzes the oxidation of lactate with concomitant reduction of NAD+ to NADH. Reductase uses NADH and reductase substrate to generate luciferin, which is converted to a bioluminescent signal by Ultra-Glo™ rLuciferase. The luminescent signal generated is proportional to the amount of LDH present.

Lactate dehydrogenase detection with LDH-Glo cytotoxicity assay 15079ma-w

"The Promega LDH-Glo™ Assay is a reliable and easy-to-follow kit to use when measuring cell death. We use this in our lab and, compared to others, it really helps us get reliable data. Overall, we would highly recommend this kit."

LDH-Glo Customer

Monitor Cytotoxicity in 3D Cell Cultures

The LDH-Glo™ Cytotoxicity Assay is well-suited for measuring LDH released from small numbers of membrane-damaged cells. Here is an example of measuring drug-induced toxicity in HCT116 spheroids. The time-dependent toxicity measurements were performed by repeatedly sampling media from the same wells.

Measuring drug-induced toxicity in HCT116 spheroids using the bioluminescent ldh assay 15086mb-wb — HCT116 spheroids were formed using 2,500 cells/well in Corning 384-well Ultra-Low Attachment plates and treated with Doxorubicin. 2.5µl samples were collected from the same wells at the indicated time points and frozen in LDH Storage Buffer at a 1:10 dilution. Samples were thawed and further diluted 2.5-fold in LDH Storage Buffer before measuring LDH levels.

See more data: Learn how this assay is used with different model systems, including SKBR3 cells to assess an antibody drug conjugate and in 3D human liver microtissues to monitor cytotoxicity over time, in the poster: New Sensitive Luminescent LDH-release Assay that Enables “Real-Time” Sampling from Individual 3D Microtissues, presented at SLAS Europe 2018.

Download Poster

Detect Target Cell Cytotoxicity in ADCC Assays

The LDH-Glo™ Cytotoxicity assay can be used to assess cell death due to many mechanisms of action of biologics treatments, including complement dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-drug conjugate cytotoxicity. In this example, the LDH-Glo™ Assay was used to detect ADCC killing of target cells using the biologic rituximab.

Detection of target cell cytotoxicity in ADCC assays using a bioluminescent ldh assay method 15087mb-wa — Cells were plated at a 20:1 Effector (PBMC):Target (Daudi) ratio and treated with varying concentrations of rituximab (RTX) in triplicate for 4 or 6 hours. Medium samples (2.5µl) from the same wells were removed and diluted into LDH Storage Buffer (47.5µl) and frozen. 25µl of thawed samples were added to an equal volume of LDH Detection Reagent and luminescence was recorded after 60 minutes incubation at room temperature. Concentration of rituximab versus RLU was plotted and data was fit to 4PL curve to calculate EC₅₀.

Detection of target cell cytotoxicity in ADCC assays using a bioluminescent ldh assay method 15087mb-wb — RLU values from control wells (performed on the same plate as the rituximab treatment) indicate that there is minimal luminescent signal from target or effector cells alone, and that addition of rituximab is required for cell killing. ADCC = Effector cells +Target cells + RTX. RLU = Relative Luminescence Units.

Protocolos

Complete Protocol

LDH-Glo™ Cytotoxicity Assay Technical ManualPDF (1983 KB) – English

Contenido

Item	Part #	Presentación
Reductase Substrate	G885A	1 × 55μl
Lactate Dehydrogenase	J195A	1 × 1 each
LDH Detection Enzyme Mix	J238A	1 × 10ml

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

Patentes y Exclusiones

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

Contenido

Item	Part #	Presentación
Reductase Substrate	G885B	1 × 275μl
Lactate Dehydrogenase	J195A	1 × 1 each
LDH Detection Enzyme Mix	J238B	1 × 50ml

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

Patentes y Exclusiones

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

Recursos

Artículos

Menciones

Cytotoxicity Assays: In Vitro Methods to Measure Dead Cells.

--
2019 Assay Guidance Manual
Ver todas las menciones

Otros

Productos Similares

CellTox™ Green Cytotoxicity Assay

Mide los cambios en la integridad de la membrana. Monitoriza cinéticamente la citotoxicidad hasta 72 horas y además tiene capacidad multiplex.

G8741, G8742, G8743, G8731

CytoTox-Glo™ Cytotoxicity Assay

Ensayo de citotoxicidad basado en luminiscencia, de alta sensibilidad que mide el número relativo de células muertas.

G9290, G9291, G9292

CytoTox-ONE™ Homogeneous Membrane Integrity Assay

Método fluorométrico homogéneo basado en la LDH que le permite cuantificar las células no viables.

G7890, G7891, G7892

CytoTox-Fluor™ Cytotoxicity Assay

Ensayo fluorescente homogéneo de adición de un solo reactivo, que le permite medir el número relativo de células muertas en poblaciones celulares.

G9260, G9261, G9262

Promega's Política de cookies

Simple, More Sensitive LDH Assay Method to Determine Cytotoxicity

Monitor Cytotoxicity from Small Numbers of Cells

How the Assay Works

Monitor Cytotoxicity in 3D Cell Cultures

Detect Target Cell Cytotoxicity in ADCC Assays

Protocolos

Complete Protocol

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Recursos

Artículos

Menciones

Otros

Promega's Política de cookies

LDH-Glo™ Cytotoxicity Assay

Simple, More Sensitive LDH Assay Method to Determine Cytotoxicity

Monitor Cytotoxicity from Small Numbers of Cells

How the Assay Works

Monitor Cytotoxicity in 3D Cell Cultures

Detect Target Cell Cytotoxicity in ADCC Assays

Protocolos

Complete Protocol

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Especificaciones

Contenido

SDS

Certificado de Análisis

Use Restrictions

Condiciones de Almacenaje

Patentes y Exclusiones

Recursos

Artículos

Menciones

Otros

Productos Similares

CellTox™ Green Cytotoxicity Assay

CytoTox-Glo™ Cytotoxicity Assay

CytoTox-ONE™ Homogeneous Membrane Integrity Assay

CytoTox-Fluor™ Cytotoxicity Assay

Usado con frecuencia junto con

CellTiter-Glo® 3D Cell Viability Assay

GloMax® Discover System

RealTime-Glo™ MT Cell Viability Assay

Caspase-Glo® 3/7 Assay System

Americas

Pacific Asia

Europe