Overview

  • Product name
    Calcium Assay kit (Fluorometric)
    See all Calcium kits
  • Detection method
    Fluorescent
  • Sample type
    Serum, Plasma, Cell Lysate
  • Assay type
    Quantitative
  • Sensitivity
    0.03 mM
  • Range
    0.04 mM - 1.5 mM
  • Assay time
    0h 30m
  • Product overview

    Calcium Assay Kit (Fluorometric) ab112115 provides a simple method for detecting calcium in physiological solutions by using a red fluorescence probe. The fluorescence signal can be easily read by a fluorescence microplate reader at Ex/Em = 540/590 nm.


    The kit can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. The calcium assay can be completed within 30 minutes.


    The calcium assay kit can be used to detect as little as 0.03 mM calcium. The kit has a broad dynamic range (30 μM to 1 mM).


    Calcium assay protocol summary:
    - add samples and standards to wells
    - add reaction mix
    - incubate for 5-30 min whilst measuring the fluorescence intensity with a microplate reader

  • Notes

    This product is intended to be used in vitro with solution-based samples such as cell extracts. If you would like to detect calcium in vivo using live cells, please check our Fluo-8, Rhod-4 or Fura-2 based assays.

    Previously called Calcium Quantification Kit.

  • Platform
    Microplate reader

Properties

Images

  • Ca measured in biologicals showing concentration (mM)

  • Standard curve (fluorimetric): mean of duplicates (+/-SD) with background readings subtracted

Protocols

References

This product has been referenced in:
  • White MM  et al. Neutrophil Membrane Cholesterol Content is a Key Factor in Cystic Fibrosis Lung Disease. EBioMedicine 23:173-184 (2017). Read more (PubMed: 28835336) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Answer

The assay buffer in ab112115 is 50 mM Na borate, pH = 8.5.

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Answer

Component B is a simple sodium borate buffer and therefore is just a dilution buffer.

The optimum pH for this dye is pH8.5. It is quite stable as long as it is protected from the light so you should be able to observe fluorescence after 24 hours without any problems.

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Answer

Thank you for contacting us.

There is always some degree of non specific binding with the fluorophore and plastic of microwell plate. We are not aware if the interaction is chemical however we recommend our customer sot use negative controls in order to deduct the background staining due to non specific binding.

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

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Answer

Thank you for contacting us.

Unfortunately we do not have any data to prove serum interferes with the dye. However, in order to overcome this problem you could add the same amount of serum as sample into the standard.

I hope this helps. Please do not hesitate to contact us for further assistance.

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Answer

Thank you for contacting us.

In regards to your questions, we would recommend freezing the stock solution instead of the reaction mix, however, there should not be a problem if it is stored at -20C and away from light for up to a week.

I hope this helps. Please do not hesitate to contact us for further assistance.

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Question
Answer

Merci de nous avoir contactés.

Malheureusement, le kit ab112115 ne devrait pas être utilisé dans un milieu acide. Un tampon acide pourrait conduire à des interférences et donc diminuer la fluorescence.

Nous suggérerons aux clients d’utiliser ce kit à un pH optimal de 8,5 avec notre tampon à 50 mM. Vous pouvez commencer avec un pH de 6,5, en utilisant le tampon à une concentration d’environ 10 mM.

Sinon, je vous recommande de regarder sur le site Web Biocompare qui vous permettra de rechercher des produits biologiques et leurs fournisseurs. Le lien est le suivant:
http://www.biocompare.com/

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Answer

Thank you for contacting us and your interest in our products.

The Calcium Quantification Assay kit (Fluorometric), ab112115, is intended for use with solution based samples. For your experiment, it may be better to have a look at the Fluo-8 Medium Removal Calcium Assay kit (ab112128) or the Fluo-8 No Wash Calcium Assay kit (ab112129) to see if you think these would be suitable for you.

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to contact us again.

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Answer

Thank you for contacting us.

I have brought your question to the attention of the lab who have informed me that you can wait a few hours with no effect.

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

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Answer

Thank you for getting back to me.

Yes, the signal would be expected to be enhanced in the presence of zinc.

And in reply to your following question, yes the Fluo-8 (ab112129) and Rhod-4 (ab112157) would not be suitable for your samples. I am sorry for the confusion caused.

If you have any further questions, please do not hesitate to ask.

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Answer

Thank you for contacting us yesterday in regards to the Calcium Quantification Assay kit (Fluorometric), ab112115.

I now have some further information to share with you in regards to the kit. Unfortunately, I can confirm that the kit is likely to cross-react with zinc (as well as cadmium, mercury, lead and manganese). I am sorry but we do not currently have any kits available from our catalogue where this is not going to be a problem for you (I am confirming this with one final kit and I will get back to you with this information as soon as I have it).

I can however say that the protein in your aqueous solution should not significantly affect the assay.

As discussed over the phone, this kit can measure down to 0.03mM. If you are interested in detecting levels much below this I would suggest having a look at the Fluo-8 No Wash Calcium Assay kit (ab112129). This kit is able to measure much lower calcium concentrations (however, this kit will also pick up zinc).

I hope this information has been of some help. If you have any further questions, please do not hesitate to contact us again.

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1-10 of 15 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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