EdU (5-ethynyl-2'-deoxyuridine)
EdU (5-ethynyl-2'-deoxyuridine)
Invitrogen™

EdU (5-ethynyl-2'-deoxyuridine)

Green features
EdU (5-ethynyl-2’-deoxyuridine) can be used as a replacement for BrdU (5-bromo-2’-deoxyuridine) and directly measures de novo DNA synthesis or S-phaseRead more
Have Questions?
Change viewbuttonViewtableView
Catalog NumberQuantity
E104155 g
A1004450 mg
E10187500 mg
Catalog number E10415
Price (USD)
6,993.65
Online Exclusive
7,210.00
Save 216.35 (3%)
Each
Add to cart
Quantity:
5 g
Price (USD)
6,993.65
Online Exclusive
7,210.00
Save 216.35 (3%)
Each
Add to cart

EdU (5-ethynyl-2’-deoxyuridine) can be used as a replacement for BrdU (5-bromo-2’-deoxyuridine) and directly measures de novo DNA synthesis or S-phase synthesis of the cell cycle using click chemistry. Click chemistry is a method of covalently coupling an azide with an alkyne. Detection of EdU employs the copper(I) catalyzed click reaction with an azide modified fluorescent dye to form a stable triazole ring. Because of the small size of the click detection reagent, no harsh denaturation steps are needed to gain access to the DNA. Eliminating this step allows for more reproducible results, a simpler and quicker protocol and measurements which can easily be multiplexed with relevant antibody based targets including phospho-histone H3, Ki-67, and cyclin B1 as well as dual-pulse experiments with BrdU by flow cytometry, fluorescence microscopy, microplate (HTS) or high-throughput imaging (HCS).

EdU is readily soluble in DMSO, alcohol, water, or aqueous buffers. For use with in vitro applications, prepare 10 mM EdU stock solution in DMSO or aqueous buffer.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
FormatSolid
Label or DyeEdU
Quantity5 g
Shipping ConditionRoom Temperature
Product LineClick-iT™
Unit SizeEach
Contents & Storage
Store at ≤-20°C, and desiccate.
PG2624-PJT8477-COL022341-Media-Promo-Pod-Graphic-Global-F-278x123
four-plex-image-COS7-live-cell zoomed

Frequently asked questions (FAQs)

What is the solubility of EdU in water? How about solubility in DMSO?

Maximum solubility of EdU is to a concentration of 25 mM in water and up to 100 mM in DMSO.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am observing no signal or very low specific signal for my click-labeled samples. What can I do to improve the signal?

The click reaction is only effective when copper is in the appropriate valency. Azides and alkynes will not react with each other without copper. Make sure that the click reaction mixture is used immediately after preparation when the copper (II) concentration is at its highest.
Do not use additive buffer that has turned yellow; it must be colorless to be active.
Cells need to be adequately fixed and permeabilized for the TdT enzyme and click reagents to have access to the nucleus. Tissue samples require digestion with proteinase K or other proteolytic enzymes for sufficient TdT access.
Some reagents can bind copper and reduce its effective concentration available to catalyze the click reaction. Do not include any metal chelator (e.g., EDTA, EGTA, citrate, etc.) in any buffer or reagent prior to the click reaction. Avoid buffers or reagents that include other metal ions that may be o xidized or reduced. It may be help to include extra wash steps on the cell or tissue sample before performing the click reaction.
You can repeat the click reaction with fresh reagents to try to improve signal. Increasing the click reaction time longer than 30 minutes will not improve a low signal. Performing a second, 30 minute incubation with fresh click reaction reagents is more effective at improving labeling.
Your cells may not be apoptotic. Prepare a DNase I-treated positive control to verify that the TdT enzymatic reaction and click labeling reaction are working correctly.

Find additional tips, troubleshooting help, and resources within our Labeling Chemistry Support Center.

I am observing high non-specific background when I image my Click-iT EdU TUNEL-labeled samples. What is causing this and what can I do to reduce the background?

The click reaction is very selective between an azide and alkyne. No other side reactions are possible in a biological system. Any non-specific background is due to non-covalent binding of the dye to various cellular components. The Select FX Signal Enhancer is not effective at reducing non-specific charge-based binding of dyes following the click reaction; we do not recommend its use with the Click-iT detection reagents. The best method to reduce background is to increase the number of BSA washes. You should always do a no-dye or no-click reaction control under the same processing and detection conditions to verify that the background is actually due to the dye and not autofluorescence. You should also perform the complete click reaction on a no-TdT enzyme control sample to verify the specificity of the click reaction signal.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I notice that when I post-stain my cells with DAPI after performing the click reaction to detect EdU incorporation, my DAPI signal is lower compared to my no-click reaction control samples. What causes the reduction in DAPI signal?

The copper in the click reaction denatures DNA to a small extent (although not as much as is required for efficient BrdU detection), which can affect the binding affinity of DNA dyes including DAPI and Hoechst stain. This effect should only be apparent with the classic EdU kits and not the Click-iT Plus EdU kits, which use a lower copper concentration.

Find additional tips, troubleshooting help, and resources within our Cell Viability, Proliferation, Cryopreservation, and Apoptosis Support Center.

I am observing no signal or very low signal for my click-labeled samples. What can I do to improve the signal?

The click reaction is only effective when copper is in the appropriate valency. Except for the DIBO alkyne-azide reaction, azides and alkynes will not react with each other without copper. Make sure that the click reaction mixture is used immediately after preparation when the copper (II) concentration is at its highest.
Do not use additive buffer that has turned yellow; it must be colorless to be active.
Cells need to be adequately fixed and permeabilized for the click reagents to have access to intracellular components that have incorporated the click substrate(s).
Some reagents can bind copper and reduce its effective concentration available to catalyze the click reaction. Do not include any metal chelator (e.g., EDTA, EGTA, citrate, etc.) in any buffer or reagent prior to the click reaction. Avoid buffers or reagents that include other metal ions that may be oxidized or reduced. It may be help to include extra wash steps on the cell or tissue sample before performing the click reaction.
You can repeat the click reaction with fresh reagents to try to improve signal. Increasing the click reaction time longer than 30 minutes will not improve a low signal. Performing a second, 30 minute incubation with fresh click reaction reagents is more effective at improving labeling.
Low signal can also be due to low incorporation of EdU, EU, or other click substrates. Other click substrates (e.g., AHA, HPG, palmitic acid, azide, etc.) incorporated into cellular components may have been lost if not adequately cross-linked in place or if the wrong fixative was used. For click substrates that are incorporated into the membrane or lipids, you should avoid the use of alcohol or acetone fixatives and permeabilizing agents.
The incorporated click substrate must be accessible at the time of the click reaction; labeling of incorporated amino acid analogs may be lower in native proteins relative to denatured proteins.
You may need to optimize the metabolic labeling conditions including analog incubation time or concentration. Cells that are healthy, not too high of a passage number and not too crowded may incorporate the analog better. You may create a positive control by including extra doses of the click substrate during multiple time points during an incubation time that spans or closely spans the doubling time of the cell type of interest.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Documents & Downloads

Certificates

Safety Data Sheets

Share catalog number, name or link

1x1 image pixel for data collection