Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Live and Dead Cell Assay (ab115347)

Submit a review Q&A (7)References (44)
Functional Studies - Live and Dead Cell Assay (ab115347)
  • Functional Studies - Live/Dead Cell Assay (ab115347)
  • Functional Studies - Live/Dead Cell Assay (ab115347)
  • Functional Studies - Live/Dead Cell Assay (ab115347)

Key features and details

  • Detection method: Fluorescent
  • Platform: Flow cytometer, Fluorescence microscope

Overview

  • Product name

    Live and Dead Cell Assay
    See all Live dead cell kits

  • Detection method

    Fluorescent

  • Product overview

    Live Dead Assay Kit ab115347 differentially labels live and dead cells with fluorescent dyes with a one-step live dead assay protocol. It is used for the rapid quantitation of cell viability using flow cytometry or fluorescent microscopy. 


    The Live Dead assay staining solution is a mixture of two fluorescent dyes that differentially label live and dead cells. 


    The Live cell dye labels intact, viable cells green. It is membrane permeant and non-fluorescent until ubiquitous intracellular esterases remove ester groups and render the molecule fluorescent. The Excitation (max) and Emission(max) are 494nm and 515nm (similar to FITC).


    The Dead cell dye labels cells with compromised plasma membranes red. It is membrane-impermeant and binds to DNA with high affinity. Once bound to DNA, the fluorescence increases >30-fold. The Excitation (max) and Emission(max) are 528nm and 617nm. 


    The Live Dead assay protocol uses a one-step staining procedure that is simple and fast. It can be used directly in cell culture media.

  • Notes

    This assay is not suitable for use with fixed cells / cell fixation.

    The Live Dead assay staining solution provided is sufficient for ~1000 assays.

    Related assays

    Review the cell health assay guide to learn about kits to perform a cell viability assay, cytotoxicity assay and cell proliferation assay. 

  • Platform

    Flow cytometer, Fluorescence microscope

Properties

Images

  • Functional Studies - Live and Dead Cell Assay (ab115347)
    Functional Studies - Live and Dead Cell Assay (ab115347)

    Dot plots showing live/dead analysis of vehicle or drug treated Jurkat cells (day 3 of treatment). The indicated drug is used to induce cell death. Live cells are on the y-axis and dead cells are on the x-axis. The red polygongate identifies live cells and the number indicates the percent of live cells.



     

  • Functional Studies - Live/Dead Cell Assay (ab115347)
    Functional Studies - Live/Dead Cell Assay (ab115347)

    Quantification of % viable cells of Jurkat cells treated with a dose response of the inidicated drug (to induce cell death) and analyzed using the live/dead assay stain on days 1, 2 and 3 using flow cytometry.

     

  • Functional Studies - Live/Dead Cell Assay (ab115347)
    Functional Studies - Live/Dead Cell Assay (ab115347)

    The sample dot plots demonstrate varying ratios of live and dead cells. More green = upper left = live cells; more red = lower right = dead cells.

  • Functional Studies - Live/Dead Cell Assay (ab115347)
    Functional Studies - Live/Dead Cell Assay (ab115347)

    Jurkat cells stained with the live/dead assay kit. Jurkat cells treated with a drug to induce cell death were labeled with the live/dead assay stain. Live cells (with esterase activity) stain green and dead cells (compromised plasma membrane) stain red. (A) Field of cells following 10 minute staining in media of live/dead stain. (B)Magnified view showing that in live cells the whole cell is stained green whereas in dead red cells it is the fragmented nuclear DNA that is stained.

References (44)

Publishing research using ab115347? Please let us know so that we can cite the reference in this datasheet.

ab115347 has been referenced in 44 publications.

  • Jorgensen M  et al. Alginate Hydrogel Microtubes for Salivary Gland Cell Organization and Cavitation. Bioengineering (Basel) 9:N/A (2022). PubMed: 35049747
  • Döhla J  et al. Metabolic determination of cell fate through selective inheritance of mitochondria. Nat Cell Biol 24:148-154 (2022). PubMed: 35165416
  • Bolanta SO  et al. Synthesis of Poly(acrylic acid)-Cysteine-Based Hydrogels with Highly Customizable Mechanical Properties for Advanced Cell Culture Applications. ACS Omega 7:9108-9117 (2022). PubMed: 35350353
  • Jia H  et al. Injectable hydrogel with nucleus pulposus-matched viscoelastic property prevents intervertebral disc degeneration. J Orthop Translat 33:162-173 (2022). PubMed: 35415072
  • Hamidi M  et al. Anionic exopolysaccharide from Cryptococcus laurentii 70766 as an alternative for alginate for biomedical hydrogels. Int J Biol Macromol 212:370-380 (2022). PubMed: 35613678
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-7 of 7 Abreviews or Q&A

Question

Product code: 115347
Inquiry: Dear Abcam Could you send me the spectra data of the green and the red stain. Or could you tell me if the channels FITC and PE from our BD CantoII FACS will be able to: 1. Measure 2. Compensate Thank you for your help, Kind regards,

Read More
Answer

Thank you for contacting us.

You can use same FITC and PE filters in this assay.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

Do you have a sample size of this Live/Dead cell assay kit?

We would like to try it with our cells.

Read More
Answer

Thank you for contacting us.

Unfortunately, we do not have free or trial sized samples of any of our products. We will guarantee this kit for differentiating between live and dead cells. If you are at all unsatisfied with the results, we are more than happy to refund or replace the kit as long as you contact us within six months of purchase. I hope this helps, please let me know if you need any additional information or assistance.

Read More

Question

Your e-mail of 30 th of may to Henriksen Stian - se below

We would like a full refund to our bank account

Read More
Answer

Thank you for contacting us.

I have sent refund request to our accounts department on 13th June 2012. If you haven't received the refund please contact them by email mailto:creditcontrol@abcam.com or call 01223-696900 for accounts.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

We are still not satisfied with the live/dead cell assay kit and we also have some comments to the reply from the developers of the product:

- Primary cells tends to be very sensitive to changes in the environment. Our cells will start to lose their original morphology in PBS and eventually round up and detach. Staining in PBS is not an option for our cells.

- In fact, we have tried to increase incubation up to 30 min without being able to see any change except that more cells detach and die because of sub-optimal conditions. Thus, increasing incubation time will give us false results (more DNA staining).

- As always when working with fluorescent microscopy, the staining procedure and stained cells are kept protected from light.

- Both the protocol and the tube with the stain say “store at 4 °C”. You also claim that the stain is stable for 6 months at 4 °C. If the stain should be stored at a different temperature you should change your protocol and labeling.

- We are not in possession of jurkat cells and are not able to test the stain with these cell line. We mainly work with primary cells.



Please, we cannot use any more time with this product and we still would like a refund for this purchase. Can you please inform us of which measures we should take to achieve this?

Read More
Answer

Thank you for contacting us.

Your credit note ID is XXXXXXX

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Question

We have now tried trypsinizing the cells and the stain works better this way. We have also tried the stain on other adherent primary cells, but the green stain is quite faint here as well.
We actually bought this kit to monitor adherent cells and their morphology. The morphology is of importance since we work with virus-infected cells. By trypsinization we also lose all detached cells. However, investigation of cell death in these cells is of great importance. From the information we found on your website and the kit protocol we were confident that this kit would be able to do this in adherent cells as well as trypsinized cells.
This stain is therefore not of much use for us and we hope that it would be possible to get a refund for this purchase. How should we proceed to accomplish this?
The order was made in January 2012 and the order number is: #1014162
Regards

Read More
Answer

Thank you for contacting us. I have been discussing this case with my colleagues, who developed this product; the following is their reply

-try staining the cells in PBS+dye (aspirate media, replace with 5X dye in PBS) [2b in microscopy section of protocol]

-it may be worth trying a staining time of longer than 10 min (which may increase the green stain)

-the green dye does have a tendency to bleach, so samples really do need to be kept in the dark and care needs to be taken in imaging to acquire images quickly before bleaching occurs.

-storage and handling of this reagent is important-should be stored at -20 and should not go through many freeze/thaw cycles. The protocol recommends aliquoting to single use vials.

- the product was quality checked using the jurkat cell line so these could be a good positive control. We never used kidney epithelial cells so we are unable to confirm the suitability of this kit. Please use Jurkat cells as positive control.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

Lot number: GR61229-2
Inquiry: Hi, We are not able to make the live/dead cell assays live cell stain work in fluorecsent microscopy. The dead cells are stained nicely and according to our expectations, but all the other cells remains unstained with only a weak green background. We have used the assay according to the protocol. The stain is diluted in PBS to twice the final concentration and then added to the cell media resulting in an additional two-fold dilution of the staining solution. The cells are then incubated at RT for 10 minutes in the dark before imaging. We have tried a final stain concentration of 5x and 10x. The cells we use are adherent primary human renal proximal tubula epithelial cells (hRPTECs) grown for 3 days in renal epithelial growth medium before staining. We use a Nikon microscope (TE2000-U) with EGFP Filter cube for TE2000 (41017), Bandpass( ex470/40,em525/50) and Chroma filterblock (41021) CY 3,5 (ex565/30, em620/60). Hope that you are able to help us with this problem. Best regards

Read More
Answer

Thank you for your email. I am sorry to hear that you are experiencing problems with this kit.

I have checked the protocol and details you have kindly provided. It seems the dye is not entering the cells, which could be due to cells so is it possible if you can try the Jurkat cells, which will be a good positive control?

Could you try trypsinizing the cells.

Le t me know if the results improves, if notI will then let lab know about this lot.

Read More

Question

Dear sir,madam, The live/dead staining kit (ab115347), does it allow to keep on culturing the living cells for a longer period of time after staining? Kind regards,

Read More
Answer

Thank you for your enquiry. I am sorry to confirm we have not tested cell viability post-staining with this product and we would not be able to guarantee whether or notit would affect the cells you want to continue culturing. Ifyou would liketo test it outyou would need to design a pilot experimentto investigate whether or not stained vs. un-stained cells have a difference in viability. Alternatively, we can suggest to simply staining a small portion of the culture for live/dead determination while retaining the remaining majority of the cells unstained in culture. This is the intended/anticipated application of this assay. I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com