Useful for staining and quantitation of the accumulated oil in adipocyte

Lipid Assay Kit


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Background

Adipocyte accumulates neutral lipid in the cell maturing and neutral lipid is accumulated as lipid droplet. The evaluation of the amount of lipid droplet in the cell becomes criterion of the differentiation of the adipose cell.

Oil red O stain is one of the methods for determination of differentiation in adipocyte. Oil red O, the oilphilic red dye, stains intracellular lipid droplet. This lipid assay kit, consisting of cell fixative and oil red O solution, is used to stain the accumulated oil in adipocyte. Furthermore, this kit is able to quantify amount of oil with organic solvent extraction method.
This kit developed as option kit of White Adipocyte Culture Kit, Brown Adipocyte Culture Kit, Visceral Adipocyte Culture Kit. Put this kit to use oil stain and quantitation in adipocyte.


Human Subcutaneous Adipocyte stained
with Lipid Assay Kit

Kit component

Component Quantity Storage
Oil O solution 150 mL x 2 4C
Solnent for red O sxtraction 200 mL x 2 4C

*One kit contains reagent for 30 x 24 well plates

Prepare reagents

EFixative solution

37 % formaldehyde 100 mL
Distilled or deionized water 900 mL
NaH2PO4(H2O) 4 g
Na2HPO4 6.5 g

 

EOil red O working solution

1. Mix Oil red O solution and distilled water in the proportion of six to four, and leave it for 10~15 minutes at room temperature.

2. Filtrate Oil red O working solution by 0.5~1.0μm syringe filter. Note: The working solution is stable for no longer than 2 hours. The solution containing crystallized Oil red O cannot be used.

Protocols (24-well plate format)

1. Remove culture medium, and wash each well once with 500 μL of PBS.

2. Add 500μL of Fixative to each well, fix it over night at room temperature.

3. After the fixation, wash each well three times with 500μL of distilled water.

4. Add 500μL of Oil red O solution to each well, and leave at least for 15min in room temperature.

5. Remove Oil red O solution, wash each well three times with distilled water. Repeat until the distilled water is perfect clean.

6. Dry it, and observe. In addition, add 500μL of extraction reagent to each well, measure dye extraction (540 nm) by spectrophotometer or plate reader.

Application Example

Viceral adipocytes (PMC-VAC01-COS) were stained by Lipid Assay Kit.
Viceral adipocytes were stained with Oil red O solution (Left figure), and then dye extractions were measured by spectrophotometer (540 nm) (Right Figure).

PMC_AK09FCOS_1

References

(1) Tashiro, K., Inamura, M., Kawabata, K., Sakurai, F., Yamanishi, K., Hayakawa, T., Mizuguchi, H. Efficient Adipocyte and Osteoblast Differentiation from Mouse Induced Pluripotent Stem Cells by Adenoviral Transduction. Stem Cells. 27, 1802-1811 (2009)

Product List

Product Name Cat# Quantity Price

Lipid Assay Kit DataSheet ,

PMC-AK09F-COS

1 SET ¥37,000
$494

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.



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