Lipase Assay Kit (Colorimetric) (ab102524)

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Overview

  • Product name

    Lipase Assay Kit (Colorimetric)
    See all Lipase kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Milk, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media

  • Assay type

    Enzyme activity

  • Sensitivity

    0.02 mU/well

  • Assay time

    1h 30m

  • Product overview

    Lipase Assay Kit ab102524 is a rapid, simple, and sensitive colorimetric assay for the measurement of lipase activity.


    In the lipase assay protocol, lipase hydrolyzes a triglyceride substrate to form glycerol which is quantified enzymatically by monitoring a linked change in the absorbance of a probe (OD=570nm). This kit is suitable for high throughput analysis (HTP).


    This lipase assay kit detects lipase activity as low as 0.02mU per well.


    Lipase assay protocol summary:
    - add samples and standards to wells
    - add reaction mix
    - analyze every 2-3 min with microplate reader in kinetic mode for at least 60-90 min at 37ºC

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

Images

  • Functional Studies - Lipase Activity Colorimetric Assay Kit (ab102524)
    Functional Studies - Lipase Activity Colorimetric Assay Kit (ab102524)Image from Mondragon A et al., PLoS One. 2014;9(8):e104873. Fig 3.; doi: 10.1371/journal.pone.0104873. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Plasma lipase levels were measured (using ab102524) after 75 days treatment with saline, liraglutide, exendin-4 or sitagliptin. ND, normal chow diet; HFD, high fat diet. p≤0.05, *; p≤0.01, **, n = 3–7 mice.

    There were no detectable differences in plasma lipase activity in mice on a normal chow diet administered any of the three drugs when compared to animals administered saline. Likewise, there was no significant change in plasma lipase activity in mice that were administered saline on a high fat diet vsnormal diet. Furthermore, administration of liraglutide and exendin-4 in combination with a high fat diet also failed to affect plasma lipase activity. We observed no detectable changes in plasma lipase activity in animals maintained on a normal chow diet and administered any of the three drugs when compared to animals administered saline.

  • Functional Studies - Lipase Activity Colorimetric Assay Kit (ab102524)
    Functional Studies - Lipase Activity Colorimetric Assay Kit (ab102524)
    Sample Timeline
  • Functional Studies - Lipase Activity Colorimetric Assay Kit (ab102524)
    Functional Studies - Lipase Activity Colorimetric Assay Kit (ab102524)
    Glycerol standard Curve

References

Publishing research using ab102524? Please let us know so that we can cite the reference in this datasheet.

ab102524 has been referenced in 4 publications.

  • Sakamuri SSVP  et al. Absence of Tissue Inhibitor of Metalloproteinase-4 (TIMP4) ameliorates high fat diet-induced obesity in mice due to defective lipid absorption. Sci Rep 7:6210 (2017). PubMed: 28740132
  • Wewer Albrechtsen NJ  et al. Glucagon-like Peptide 1 Receptor Signaling in Acinar Cells Causes Growth-Dependent Release of Pancreatic Enzymes. Cell Rep 17:2845-2856 (2016). PubMed: 27974199
  • Ma X  et al. The oncogenic microRNA miR-21 promotes regulated necrosis in mice. Nat Commun 6:7151 (2015). PubMed: 25990308
  • Mondragon A  et al. Divergent effects of liraglutide, exendin-4, and sitagliptin on beta-cell mass and indicators of pancreatitis in a mouse model of hyperglycaemia. PLoS One 9:e104873 (2014). PubMed: 25119717

Customer reviews and Q&As

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1-10 of 14 Abreviews or Q&A

Question

can kit be used with vegetable oil (purified)?
Has this been tried?

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Answer


Since the lab has not tried vegetable oil themselves, they do not have any recommendations/ comments on what the most optimal protocol for such use would be. That would have to be optimized by the end user.

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Question

I'm using this kit, but my samples are showing a weak signal. I'm using rat serum previously frozen at -80C for 4 months. Do you think the lipase could have degraded in this time? Any recommendations to increase the signal?

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Answer

Thank you for your inquiry.

I heard back from the lab that the efficiency of the assay depends on how well the serum was frozen, how many times it has been freeze thawed since then etc.

If the serum is of good quality, the lab suggests starting with more amounts of it to get signals within the linear range of the standard curve. Did you follow the Calcium addition recommendation for specific lipases? Some Lipases require calcium and if your lipase requires calcium you should avoid EGTA in sample preparation and add calcium (1-5 mM) to the Lipase assay buffer before use.
The two samples shown in the datasheet are different amounts of the positive control. Did the positive control work well?
I hope this information has been useful for you. Please let me know if you have any other questions.

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Question

Can the Assay Buffer provided with the kit be used with a biomasher type mortar and pestle to produce a cell extract?

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Answer

the Assay Buffer included with the kit can be used to open cells, thus producing a homogenate that can be assed for lipase activity.

It is recommended to use a dounce homogenizer to homogenize cells or tissues here, although your biomasher may work as well. You can check to progress of cell lysis by viewing cells under a microscope.

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Question

I have one more question. Does that mean ab102524 is not a triglyceride derivative?

Thanks,

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Answer

Thank you for contacting Abcam.

The substrate for ab102524 is also a triglyceride derivative. The datasheet product overview states, "In Abcam's Lipase Assay Kit (Colorimetric), lipase hydrolyzes a triglyceride substrate to form glycerol which is quantified enzymatically by via monitoring a linked change in the OxiRed probe absorbance (?=570nm)."

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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Question

kit component: glycerol standard had only 20 ul instead of 200 ul
kit works well
lot GR91965-2

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Answer

Thank you for your patience.

I am sorry this kit component did not conatin the amount as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx. The estimated delivery date is xxx.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

kit component: glycerol standard had only 20 ul instead of 200 ul
kit works well
lot GR91965-2

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Answer

Thank you for contacting us and for letting us know about this issue.

I am in touch with the supplying lab who has agreed to provide another vial of the glycerol standard as well as with our internal department to arrange for this.

Once I have more definite information I will let you know.

I apologize for this mistake on our end and hope to resolve this issue very soon.
In the meantime, please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Question

There is still a bit of confusion regarding the method. Is the OD that you use to plot the standard curve the delta OD (i.e. T1-T2) or just OD. If it is just OD, what time should the OD be read? I have included both sets of data.
Time 10 mins
Glycerol (nmol/per well)
T1 OD570nm

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Answer

Thank you for your email.

I have received the following email form lab;

For the standards, you have to take the OD only at the final time point. The OD values of the various concentrations of glycerol will increase with time and finally plateau. If you take a reading any time after the 60 min incubation, you will get this value as the OD. A

In the samples we are taking 2 readings at T1 and T2 to account for the total lipase activity between that time. The standard curve is not a lipase activity curve; it is a simple glycerol standard curve, which is based on different substrate amounts but the same activity.

So take the OD only at the final time point for the standard curve.

Also I noticed that your client is subtracting the OD of the control from all standard values. That is incorrect. The control OD needs to be subtracted only from the sample ODs. Subtract the “0” standard value from all standard ODs.

I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

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Question

Thanks for that information – see curve below.

Thanks again

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Answer

Thank you for sending the data.

Unfortunately the graph only, is not enough for the data analysis. We need to check all the protocol details e.g. species, type of samples used, sample dilutions and the raw data etc.

I just had chat with my colleague regarding the standard curve. The OD readings seems very high; the OD should not be more than 2 for 10nmol/per well so either the glycerol standard used was very dense (not properly diluted) or the setting of machine is not correct. Could you please check this?

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Question

Hi,

Thanks for your reply. When I tried to ring you yesterday is took a long while to get through to what sounded like a engaged tone and then it sounded like the call was answered but the line was silent.

I rang again this morning to see if I could get through and I got straight through to the switch board.

Thanks again

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Answer

Thank you for letting me know.

I don't know what could have happened there but if you have any similar problems in the future please do let us know.

May I ask, when you called this morning, did you speak to any of our Scientific Support specialists? I just want to make sure that we don't come back to you with the same answers again.

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Question

Hello again,

I thought that I would email the details of the technical issue in case it is not possible to make contact via telephone. I am confused as to the protocol for the reaction mix. It states to add 100µl of sample reaction mix (assay buffer, OxiRed probe, enzyme mix & lipase substrate) to each well containing the glycerol standards, lipase positive control, and test samples. It then instructs you to add 100µl of the control reaction mix (assay buffer, OxiRed probe & enzyme mix) to each well. Is this correct? What is the enzyme mix? Shouldn’t the only enzyme be that of the sample? Why do we want to add the control to all of the samples/positive controls/standards? Isn’t the point of the standard curve to just create a calibration curve - why would you want to add sample reaction mix to it?

Thanks again

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Answer

Thank you for contacting us and sorry for the delay in getting back to you.

I am sorry for the confusion caused by how the protocol has been worded for the Lipase Detection Kit (Colorimetric), ab102524. I can confirm that as the protocol states, 100 uL of sample reaction mix should be added to each of the wells containing the glycerol standards, lipase positive control and test samples. The control reaction mix (100 uL) only needs to be added to wells containing sample controls. I.e., this allows you to measure the background signal from your samples without the lipase substrate being present.

I hope this has clarified the protocol sufficiently for you. If you have any further questions please do let me know. I will have the protocol amended to reduce this confusion in the future. I thank you for bringing this to our attention.

I am concerned that you were not able to reach us by phone yesterday. Was there any answer when you dialed our number (0122369600)? You should come through to a switch board. Did you reach this?

I look forward to receiving your reply.

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