Product manual
Product use, preparation, operating conditions and storage.
EdU proliferation detectionReagent kit
EdU-488 Cell Proliferation Detection Kit
Cat. No.QH-16-1001
Detect newly synthesized DNA through EdU incorporation and fluorescent click chemistry for proliferation and S-phase studies.
Packs & quoteEdU pulse
Fix and permeabilize
Click detection
Excitation495 nm
Emission519 nm
TargetDNA synthesized during the labeling pulse
Preparation and storage
- Storage
- Store at −20°C. Protect the fluorophore and Hoechst 33342 from light. Kit shelf life: 12 months.
EdU incorporation and CuAAC detection
EdU is a terminal-alkyne thymidine analogue incorporated into replicating DNA and detected with a fluorescent azide under copper catalysis. This kit detects after fixation and permeabilization without DNA denaturation. EdU positivity reports DNA synthesis during the pulse.
Cell-proliferation comparison
Compare treated and control groups at matched pulse times and cell conditions, alongside cell count or viability.
Microscopy and flow readouts
Microscopy retains spatial information; flow cytometry uses single-cell suspensions. Tissue sections do not directly use a suspension flow protocol.
Specifications
Product information
- Product name
- EdU-488 Cell Proliferation Detection Kit
- Cat. No.
- QH-16-1001
- Product form
- EdU proliferation detection
- Target
- DNA synthesized during the labeling pulse
- Pack sizes
- 50-500T / 200-2000T
Preparation and properties
- Excitation wavelength
- 495 nm
- Emission wavelength
- 519 nm
- Molar extinction coefficient
- 73,000 M⁻¹ cm⁻¹
- Quantum yield
- 0.92
- Storage
- Store at −20°C. Protect the fluorophore and Hoechst 33342 from light. Kit shelf life: 12 months.
- Incorporation reagent
- EdU
- Detection chemistry
- Copper-catalyzed azide–alkyne cycloaddition (CuAAC)
- DNA denaturation
- Not required
Spectral information and detection
Product parameters
Use parameters for the exact catalog number when preparing reagents, setting reagent input or calculating results. Request a lot-specific CoA for test results.
Protocol
Choose the pulse window
Set EdU concentration and pulse duration for the cell type and question, matching comparison groups.
Prepare detection components
Reconstitute for the purchased pack and prepare fresh click solution. Check staining order and compatibility with antibodies, fluorescent proteins and other dyes.
EdU pulse
Add EdU under suitable culture conditions. A reference starting concentration for A549, HeLa and NIH3T3 is 10 µM; establish pulse duration for the model.
Fix and permeabilize
Wash, fix and permeabilize using a suitable procedure. The provided fixation example is 4% paraformaldehyde for 15 minutes at room temperature; check compatibility with other markers.
Click detection
Prepare fresh reaction mixture, add components in the kit-defined order, incubate protected from light and wash thoroughly.
Acquire and quantify
Set thresholds with a no-EdU control and report positive cells, total cells and pulse duration. Include single stains for multicolor detection.
Request product documentation
Select a document and enter your phone and email. Include the lot number for a CoA.
EdU-488 Cell Proliferation Detection KitQH-16-1001
Spectra and measurement conditions
Spectra, measurement conditions and detection settings for the exact catalog number.
Certificate of analysis (CoA)
Lot-specific test results. Include the lot number from the vial.
Safety data sheet (SDS)
Handling, storage and emergency response information.
Quality and lot information
No-EdU control
Omit EdU but complete fixation, permeabilization and click detection to assess reagent background.
Known proliferating sample
Checks incorporation and click detection. Add viability assessment to investigate apparently low proliferation caused by cell damage.
Matched pulse conditions
Match EdU concentration, time, cell density and acquisition rules across comparison groups.
References
A chemical method for fast and sensitive detection of DNA synthesis in vivo
Salic A, Mitchison TJ. · PNAS · 105(7):2415–2420 · 2008
EdU incorporation is followed by fixation, permeabilization and fluorescent-azide CuAAC detection without BrdU-style DNA denaturation. EdU’s terminal alkyne is not a conventional copper-free SPAAC partner; the paper does not validate Qihe kit compatibility with GFP or PE.
Copper-Catalyzed Azide–Alkyne Click Chemistry for Bioconjugation
Presolski SI, Hong VP, Finn MG. · Current Protocols in Chemical Biology · 3(4):153–162 · 2011
Describes CuAAC catalyst systems and reaction assessment for terminal-alkyne substrates.
Conventional CuAAC EdU detection and fluorescence compatibility
Thermo Fisher Scientific · Click-iT EdU Flow Cytometry Assay Kits · MP10418, 2011
Explains the effects of copper-catalyzed EdU detection on GFP, PE and PE tandems and the importance of staining order. This is conventional Click-iT guidance; its formulation, doses and compatibility conclusions for other labels do not replace Qihe-kit validation or imply Plus-series compatibility.
Frequently asked questions
Does EdU positivity equal the entire proliferating fraction?
It reports DNA synthesis during the pulse and depends on pulse duration and cell-cycle distribution.
Does this kit detect live cells?
EdU incorporation occurs in living cells; this kit’s fluorescent detection is performed after fixation and permeabilization.
Can different colors use the same channel?
Configure channels from the SKU’s excitation/emission values and instrument filters, checking multicolor spillover.
Can detection be changed to copper-free click chemistry?
EdU carries a terminal alkyne and this kit uses CuAAC. Copper-free DBCO–azide SPAAC conditions do not replace this copper-catalyzed detection.
Can it be combined directly with GFP, PE or tandem labels?
Copper-catalyzed detection can affect GFP, PE and PE tandems. Check staining order for the purchased kit and validate single stains through fixation, permeabilization and click detection. Copper-Safe/Plus claims from other products do not apply to this kit.