Phospho & Total NDRG1 Assay Kit

Phospho-NDRG1 (Thr346), Phospho-NDRG1 (Ser330) and Total NDRG1 Cellular Assay Kit

Cisbio's cell-based homogeneous HTRF® phospho-NDRG1 assay enables quantitative detection of the modulation of NDRG1 phosphorylated on Serine 330 or Threonine 346. NDRG1 is upregulated by HIF1-alpha under hypoxic conditions often associated with aggressive tumor progression. NDRG1 is a metastasis suppressor that works by blocking the angiogenesis signaling pathway, making Cisbio’s phospho-NDRG1 assay kit a valuable tool in oncology and angiogenesis research. The phospho-NDRG1 assay can be used from basic research up to preclinical drug discovery phases due to its rapid and convenient streamlined protocol without any wash steps.  It is also amenable to low sample volume formats. This kit contains all the reagents you need, and offers increased throughput compared to ELISA or Western Blot.

   Check our lysis buffer compatibility      Check our species compatibility

Assay Principle

HTRF® - the homogeneous cell-based sandwich immunoassay

Cisbio Bioassay’s phospho- and total NDRG1 assays are based on a TR-FRET sandwich immunoassay format comprising two specific anti-NDRG1 antibodies, one labeled with Eu3+-cryptate (donor) and the other with d2 (acceptor). The antibodies specifically bind with NDRG1, and the proximity of donor and acceptor will then lead to a fluorescent TR-FRET signal. The protocol is optimized for a 384-well plate format, but can easily be further miniaturized or upscaled. Only low sample volumes are needed. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. Washing is not needed at any step.The assay can be run with frozen cell lysates or fresh cells in culture. After cell lysis, endogenous phospho- or total-NDRG1 can be quantitatively detected using the HTRF phospho-NDRG1 (Ser330), phospho-NDRG1 (Thr346) and total NDRG1 cellular assay kit reagents and most TR-FRET multimode plate readers.

A simpler, more flexible assay protocol - adapted to your applications

Two-plate assay protocol:

For added flexibility, the assay can be run as a two-plate assay protocol, where the cells are plated and treated in a 96-well culture plate. For detection, lysates are subsequently transferred to a 384sv assay plate where the HTRF® reagents are added. This also enables monitoring of the cells' viability and confluence in an appropriate cell culture plate.

Product Performance

Detection phospho and total NDRG1 on HeLa cells

Different cell densities (100K, 50K, 25K and 12.5K) of HeLa cells were plated under 50µL in 96-well plate and incubated overnight. Cells were stimulated with increasing concentration of 25 µL of AZD8055 (a novel ATP-competitive mTOR inhibitor) for 2h  at 37°C - 5% CO2. After stimulation, cells were lysed with 25µL of lysis buffer 4X for 30 min at RT under gentle shaking. 16µL of lysate were transferred into a 384-well sv white microplate and 4 µL of the HTRF phospho NDRG1 (Ser330), phospho NDRG1 (Thr346) or total NDRG1 detection reagents were added. The HTRF signal was recorded after a overnight incubation at room temperature.

Detection phospho and total NDRG1 on HeLa cells

Different cell densities (100K, 50K, 25K and 12.5K) of HeLa cells were plated under 50µL in 96-well plate in presence of 40 µM of Dp44mT and incubated for 24h. Then, media was aspirated and cells were lysed with 50µL of lysis buffer 1X for 30 min at RT under gentle shaking. 16µL of lysate were transferred into a 384-well sv white microplate and 4 µL of the HTRF phospho NDRG1 (Ser330), phospho NDRG1 (Thr346) or total NDRG1 detection reagents were added. The HTRF signal was recorded after a overnight incubation at room temperature.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
NDRG1 phospho-T346 kit - 500 tests63ADK092PEG
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NDRG1 phospho-T346 kit - 10,000 tests63ADK092PEH
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NDRG1 phospho-T346 kit - 50,000 tests63ADK092PEY
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NDRG1 phospho-S330 kit - 500 tests63ADK093PEG
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NDRG1 phospho-S330 kit - 10,000 tests63ADK093PEH
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NDRG1 phospho-S330 kit - 50,000 tests63ADK093PEY
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NDRG1 total kit - 500 tests63ADK094PEG
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NDRG1 total kit - 10,000 tests63ADK094PEH
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NDRG1 total kit - 50,000 tests63ADK094PEY
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Companion products

DescriptionCat. noProduct insertMSDS
NDRG1 phospho-T346 kit - control lysate63ADK092TDA
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NDRG1 phospho-S330 kit - control lysate63ADK093TDA
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NDRG1 total kit - control lysate63ADK094TDA
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