製品の概要

  • 製品名

    Lipase Assay Kit (Colorimetric)
    Lipase キット 製品一覧
  • 検出方法

    Colorimetric
  • サンプルの種類

    Cell culture supernatant, Milk, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media
  • アッセイタイプ

    Enzyme activity
  • 検出感度

    0.02 mU/well
  • 全工程の試験時間

    1h 30m
  • 製品の概要

    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

  • 試験プラットフォーム

    Microplate reader

製品の特性

画像

  • 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.

  • Sample Timeline
  • Glycerol standard Curve

プロトコール

参考文献

This product has been referenced in:

  • 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). Read more (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). Read more (PubMed: 27974199) »
See all 4 Publications for this product

レビューと Q&A

1-10 of 14 Abreviews or Q&A

Question
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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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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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
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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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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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!
https://www.abcam.com/abreviews

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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
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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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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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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1-10 of 14 Abreviews or Q&A

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