Overview

  • Product name

    Carboxylesterase 1 (CES1) Specific Activity Assay Kit
  • Detection method

    Colorimetric
  • Sample type

    Cell culture extracts, Tissue
  • Assay type

    Enzyme activity
  • Species reactivity

    Reacts with: Rat, Human
  • Product overview

    ab109717 (MS789) is a novel assay that uses a high affinity monoclonal capture antibody to selectively isolate carboxylesterase 1 (CES1) from liver tissue or cell samples. After isolation and subsequent measurement of the enzyme's functional activity, the quantity of isolated CES1 is measured in the same well by adding a second monoclonal detector antibody, which is quantified using a colorimetric label (HRP). Both reactions take place in time-dependent manners proportional to the amount of enzyme captured in each well. By combining activity and quantity measurements, the enzyme's relative specific activity can be determined. Specific activity is useful for measuring up or down regulation of activity by site-specific modification or damage, and in response to specific inhibitors.

     

  • Notes

    All components are shipped cold. Store all components at 4°C.

  • Platform

    Microplate reader

Properties

Images

  • Figure 1. Using this assay the acetylcholinesterase inhibitor tacrine, previosuly shown to co-crystalize in the active site of CES1, does not inhibit the activity of the enzyme which is in agreement with the crystalization study (Bencharit et al. Chem Biol 2003; 10(4) 341-9). Troglitazone a member of the thiazolidinediones class of antidiabetic drugs, was determined to inhihibit CES1 (IC50 3µM) while other members of this class had little/no effect (Fukami et al. Drug Metab Dispos 2010; 38(12):2173-8.
  • Principle of Sandwich ELISA used in Carboxylesterase 1 (CES1) Specific Activity Assay Kit (ab109717). Substrate plus HRP or AP conjugated to streptavidin provide a colorimetric signal that is readable by any plate readers capable of standard ELISA absorbance measurements.

Protocols

References

ab109717 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you very much for your call yesterday and for sending all of this data.
The kit developers at MitoSciences have reviewed the data and agree that it looks good, and the top three points can be removed to improve the fit of the regression line. There may be subtle differences between the HepG2 cells used, or in the protocol, which resulted in the different values on the standard curve, but we all agree that the data looks good.
I hope that this is useful, but if you have any further questions or if there is anything else that we can do for you, please let me know and we'll be happy to help. Have a nice day!

Read More

Answer

Thank you for your reply.

I expect the assay will still give good results with your settings, but if you do see any unexpected data, please let me know and I will be happy to help you further.

Read More

Answer

Thank you for your phone call.

My colleague expressed concerns that since the speed of your shaker is so fast, you may see your samples spread too high up the side walls, sill out of the wells, or froth. If none of these are observed, then it is probably okay. He would instead recommend using a plate rocker for the incubations or even skipping the shaking. The more important point is to ensure that there is good mixing at the development / pate reader stage.

I hope this helps, please let me know if you need any additional information or assistance.

Read More

Answer

Thank you very much for your call this morning and for your patience while I've been in touch with the scientists in the MitoSciences lab regarding this kit.

I went ahead and forwardedyour otherquestions as well, and here is the information that I've received from the lab:

The 1x bufferscan be stored at 4C for a couple of weeks, and for longer storage we would recommend freezing and thawing to room temperature before use. The strips should be resealed in the bag and stored at 4C.
For shaking, we use a 400 RPM setting. The length of shaking between kinetic readings depends on the frequency of the readings. It would be best to shake as much as possible.You can measure as frequently as your equipment can provide, and there is no loss in data for more frequent readings.We do every 15 seconds for the entire plate, but every minute would not be a problem.
Apath-length correction can help give more accurate measurements, so yes it would be better to incorporate this.
I hope that this information will be useful, but if you have any further questions or need anything else, please let me know and I'll be happy to help.

Read More

Answer

I looked at the spectrum - 405nm is the peak absorbance, but 360nm should work particularly with the wide window indicated. This assay will visibly turn yellow, so if there is any question about whether instrumentation is working correctly it is always advisable to look at the plate and check if the numbers indicated are consistent by eye I hope this is helpful. Please contact me again if you have any further questions.

Read More

Answer

Thank you for your inquiry. The protocol says to measure the wavelength at 402nm every minute for up to 1 hour for a kinetic assay. But if you would like to make an endpoint reading rather than kinetic, this can be done by observing the yellow color development in the plate and measuring it at a user-defined time. This usually happens between 10 minutes and 1 hour. In practice this is a very sensitive and linear assay, therefore you could measure the activity after 20 minutes and sufficient color has developed in the sample containing wells. I hope this information helps. Please contact us with any other questions.

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

Sign up