05 Cell Staining

Lipid Droplet Assay Kit - Blue

Lipid Droplet Assay Kit - Blue

Lipid Droplet Assay

  • High selectivity to lipid droplet (low background)
  • 2 colors available (blue and deep red)
  • Applicable to both live and fixed cell
  • Product code
    LD05  Lipid Droplet Assay Kit - Blue
Unit size Price Item Code
1 set * $256.00 LD05-10

* The typical number of usable assays : 96 well plate x 1, 40 assays for flow cytometry

ORDER
Component
1 set * ・Staining Dye - Blue
・Loading Buffer (10x)
×1
6 ml ×1

Description

Product Description
Lipi probes are small molecule which emit strong fluorescence in hydrophobic environment such as in LDs.

 

Manual

Technical info

Lipid droplets (LDs) are composed of neutral lipids such as triacylglycerol & cholesteryl ester that are surrounded by phospholipid monolayers and are seen ubiquitously, not only in adipocytes1). Although LDs were simply thought to serve as a lipid storage unit, a recent study has stated that LDs play an important role in regulating lipid metabolism, autophagy2) and cellular senescence3).Therefore, have gained great attention as an important tool to elucidate the mechanisms of formation, growth, fusion, and retraction of LDs.

1) T. Fujimoto et al., “Lipid droplets: a classic organelle with new outfits.” Histochem Cell Biol., 2008, 130(2), 263.
2) R. Singh et al., “Autophagy regulates lipid metabolism.” Nature, 2009, 458(7242), 1131.
3) M. Yokoyama et al., “Inhibition of endothelial p53 improves metabolic abnormalities related to dietary obesity.” Cell Reports, 2014, 7(5), 1691.

Only by the addition of a reagent, the imaging of lipid droplets (LDs) or the quantitative variation of LDs in live and fixed cells becomes quantifiable.

 

Lipid Droplet Assay Kit considerably shortens the entire process and can be used for live cells.
The fluorescent dye provided in the Lipid Droplet Assay Kit can be used for live and fixed cells. Compared to a method of using a colorimetric reagent, the method of using the Lipid Droplet Assay Kit can shorten measuring time. Furthermore, the repeatability of experiment can be increased by using the Lipid Droplet Assay Kit because the dye is not deposited in a plate.

Experimental example of plate assay

Changes in lipid droplets were examined after the addition of oleic acid or Triacsin C (acyl-CoA synthetase inhibitor) to the A549 cell culture medium.

As a result, we confirmed that the oleic acid-treated cells show an increase in the number of LDs, compared to control and Triacsin C-treated cells.

Blue   :Ex. 376 – 386 nm / Em 435 – 455 nm
Deep Red :Ex. 623 – 633 nm / Em 649 – 669 nm

 

Reagent Comparison

Changes in lipid droplets were examined after the addition of oleic acid or Triacsin C (acyl-CoA synthetase inhibitor) to the HeLa cell culture medium.
As a result, we confirmed that the oleic acid-treated cells show an increase in the number of LDs, compared to control and Triacsin C-treated cells.


Blue   :Ex. 405 nm/ Em 425 – 475 nm
Deep Red :Ex. 640 nm/ Em 650 – 670 nm

Related Product Information

Function     Product Size Product Code

Imaging

Lipi-Blue 10 nmol LD01
Lipi-Green 10 nmol LD02
Lipi-Red 100 nmol LD03
Lipi-Deep Red 10 nmol LD04

Quantification (Plate Reader, FCM)

Lipid Droplet Assay Kit - Blue 1 set LD05
Lipid Droplet Assay Kit - Deep Red 1 set LD06

Q & A

Q

Preparing a stock solution of oleic acid

A

Required Reagents:
・BSA (bovine serum albumin)
・Oleic acid
・0.1 mol/L Tris-HCl (pH 8.0)
Procedure:
(1) Dissolve 0.14 g/mL BSA in 0.1 mol/L Tris-HCl (pH 8.0).
(2) Add 4 mmol/L oleic acid to a disposable centrifuge tube.
(3) Add BSA solution (prepared in step 1).
(4) Cap the tube and mix on a rotary shaker (Be sure the solution is transparent, indicating that oleic acid has been conjugated to BSA).
(4) Filter the solution prepared above (step 4) using 0.22μm filter membranes.
(5) Store oleic acid stock solution at 4˚C.
*Use the appropriate amount of oleic acid stock solution for culture medium to prepare working solution.
*Oleic acid working solution cannot be stored. Please prepare the working solution immediately before usage.

Inducing lipid droplets
(1) Incubate cells for 24 hours at 37˚C in a 5% CO2 atmosphere.
(2) Add 200 µmol/L working solution (prepared from oleic acid stock solution) to culture medium and incubate for further 24 hours.

Handling and storage condition

Handling and storage condition
0-5°C
Contact
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Product Classification

Product Classification

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