Adipogenesis Detection Assay Kit (Colorimetric/Fluorometric) (ab102513)

Submit a review Q&A (8)References (7)

製品の概要

  • 製品名

    Adipogenesis Detection Assay Kit (Colorimetric/Fluorometric)
    Adipogenesis キット 製品一覧

  • 検出方法

    Colorimetric/Fluorometric

  • サンプルの種類

    Tissue, Adherent cells, Suspension cells

  • アッセイタイプ

    Quantitative

  • 検出感度

    > 0.2 nM

  • 検出範囲

    0.2 nM - 10 nM

  • 全工程の試験時間

    0h 40m

  • 製品の概要

    Adipogenesis Detection Assay Kit (Colorimetric/Fluorometric) (ab102513) quantifies triglyceride accumulation in cells and tissues. In the assay, triglycerides are efficiently solubilized then hydrolyzed to glycerol which is subsequently oxidized to convert the probe to generate color (ODmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit can detect 0.2 - 10 nmol of triglyceride in <1,000 differentiated 3T3-L1 cells. The high detection sensitivity and the convenient microplate assay format make the kit a convenient tool for studying the effect of adipogenesis inducers or to screen inhibitor compounds.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

    Adipogenesis is the process of differentiation of different cell types into adipocytes, the primary fat storage cell type. The accumulation of adipocytes is the basis for obesity, a significant risk factor in many diseases, including diabetes, atherosclerosis, cancer and cardiovascular disease, etc. Adipocytes accumulate triglycerides, in the form of lipid droplets which can be measured.

    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.

  • 試験プラットフォーム

    Microplate reader

製品の特性

画像

  • Functional Studies - Adipogenesis Detection Kit (ab102513)
    Functional Studies - Adipogenesis Detection Kit (ab102513)

    Example of a standard curve obtained with ab102513.

  • Functional Studies - Adipogenesis Detection Kit (ab102513)
    Functional Studies - Adipogenesis Detection Kit (ab102513)

    Example of a standard curve obtained with ab102513.

  • Functional Studies - Adipogenesis Detection Kit (ab102513) - Fluorometric
    Functional Studies - Adipogenesis Detection Kit (ab102513) - Fluorometric

    Glycerol measured fluorometrically in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample

  • Functional Studies - Adipogenesis Detection Kit (ab102513) - Colourimetric
    Functional Studies - Adipogenesis Detection Kit (ab102513) - Colourimetric

    Glycerol measured colourimetrically in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample.

    Ms brain value retrieved from 1:9 dilution of original 12.4mg/mL sample.

    Ms muscle value retrieved from 1:9 dilution of original 7.5mg/mL sample.

    Ms heart value retrieved from 1:27 dilution of original 16.4mg/mL sample.

  • Functional Studies - Adipogenesis Detection Kit (ab102513) - Colourimetric
    Functional Studies - Adipogenesis Detection Kit (ab102513) - Colourimetric

    Glycerol measured colourimetrically in cell lysates showing quantity (nmol) per 1 mln of tested cells

参考文献

ab102513 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab102513 は 7 報の論文で使用されています。

  • Jung YJ  et al. Cell reprogramming using extracellular vesicles from differentiating stem cells into white/beige adipocytes. Sci Adv 6:eaay6721 (2020). PubMed: 32232152
  • Xu H  et al. A long-term maternal diet intervention is necessary to avoid the obesogenic effect of maternal high-fat diet in the offspring. J Nutr Biochem 62:210-220 (2018). PubMed: 30316166
  • Li Y  et al. Suppression of adipocyte differentiation and lipid accumulation by stearidonic acid (SDA) in 3T3-L1 cells. Lipids Health Dis 16:181 (2017). Functional Studies ; Mouse . PubMed: 28946872
  • Leow SC  et al. The transcription factor SOX6 contributes to the developmental origins of obesity by promoting adipogenesis. Development 143:950-61 (2016). PubMed: 26893351
  • Llobet L  et al. OXPHOS xenobiotics alter adipogenic differentiation at concentrations found in human blood. Dis Model Mech N/A:N/A (2015). Functional Studies ; Human . PubMed: 26398948
  • Osiecki MJ  et al. Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells. PLoS One 10:e0144941 (2015). PubMed: 26660475
  • Joseph R  et al. ACSL1 Is Associated With Fetal Programming of Insulin Sensitivity and Cellular Lipid Content. Mol Endocrinol 29:909-20 (2015). PubMed: 25915184

レビューと Q&A

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1-8 of 8 Abreviews or Q&A

Question

Which type of plates can be used?

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Answer


I am happy to confirm that tissue culture plates can sustain 90°C and can therefore be used without problem in this experiment.



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Question


I work with 3T3-L1 cells, but in 12- or 6 well plates. At completion of my experiment, with culturing and treatment, should I harvest the cells, spin down, remove the supernatant and then add the 100 µL Lipid Extraction Buffer? Following which I heat and mix as in 1a iv. and v. (page 7), and then centrifuge to remove debris?

Although I have not counted, I am pretty sure a 6-well plate will give me more than 10.000 cells per well. If I plan to perform the triglyceride assay, should I then transfer 5 µL of my lipid extracts (from cells grown in 6- or 12-well plates) to a 96-well plate, or would you think this still exceeds the standard curve?

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Answer



In our hands, ˜1,000-10,000 differentiated 3T3 cells using 100ul Lipid Extraction Solution are sufficient for the colorimetric assay. Then we use 5-50ul of the extract for the assay. I would start with 5ul of the lipid extract for the assay. If this is too much, then I would do a pilot experiment using several dilutions to see which one gives me the best results within the linear range of the std. curve.


The protocol you mention is correct:

After culturing and treating your cells in a 6 or 12 well plate, you should remove the medium, wash with PBS, add 100ul lipid extraction buffer, heat for 30 mins, cool to RT and then use the liquid for assay. You can spin down to remove any debris.

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Question

I have a few questions about this kit protocol:
1. Does it matter how the lipid extraction solution is thawed? Room temp? 37 deg C water bath? One ice?
2. Is the standard available separately? I have used the standard in triplicate in my first experiment and am running low.
3. To normalize with total protein, can the lipid extract (using lipid extractionbuffer)be used as the same for runing the BCA assay or do I need to do a separate protein extraction using RIPA to measure total protein?

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Answer

The lipid extraction buffer can be thawed at RT.

Ibelieve the standard can be made available separately fromthe kit. If it is these things usually take 1-2 weeks to become available for purchase. And since itis a new product it would take 1-2 weeks after that for us to acquire stock and ship to you. Please let me know if you are interested.
To normalize with total protein, please use buffers like RIPA for lysis.
Hope this information has been helpful for you. Please let me know if you have any other questions.

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Question


Thanks for your reply.
have a good weekend!

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Answer

I'm happy to help! Please let me know if you need anything further.

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Question

Does Abcam have a customer conjugation service?

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Answer

Thank you for your enquiry.
At this time custom conjugation services are not available.
Sorry I could not be more helpful at this time. Please contact me again if you have any further questions.

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Question

What is the difference between K622-100 and K610-100?

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Answer

ab102513
Sample types: Cell and Tissue culture supernatants
Lipid extraction performed: The high detection sensitivity and the convenient microplate assay format make the kit a convenient tool for studying the effect of adipogenesis inducers or inhibitors, or to screen drugs.
ab65336:
Samples: Cell and Tissue culture supernatants, urine, plasma and serum, as well as many other biological fluids
No specific lipid extraction.
I hope this is helpful. Please contact me again if you have any further questions.

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Question

What is the sample volume and how do you normalize the data?

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Answer

Thank you for calling Abcam.

The sample volume number that you use is the volume that you originally added to the reaction at stage 1 (add 5-50ul of sample). To normalize the data, you can either divide the value you get by the number of cells you started out with or the amount of tissue.

If there is anything else I can help you with, please let me know.

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Question

This protocol is unclear. I have wasted sample.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry about the problems caused by this product. It is a good rule of thumb to always thoroughly read the protocols and call us if you have any questions at all. I would also remind you that this product only has a 6 month guarantee and shelf life. As requested, I have issued a free of charge replacement with the order number XXXX. 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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