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useful for double staining of the fat globule and the nucleus in the cell.

Adipocyte Fluorescent Staining kit

Background

The Adipocyte Fluorescent Staining Kit (Cat.No.PMC-AK19F-COS) is designed to stain both lipid droplets and nuclei in adipocytes using BODIPY? (Invitrogen Corporation) and H33258, respectively. Furthermore, the amount of lipid per cell and cell shape can be quantified using imaging instrument such as IN Cell Analyzer 1000 (GE Healthcare Company)


Human Subcutaneous Adipocyte stained with Adipocyte Fluorescent Staining kit
Lipid droplets stained with the lipophilic dye BODIPYR. The blue nuclear stain is the DNA dye Hoechst 33258,

Kit component

Component Quantity Storage
Tablet for washing 5 tablets 4-10°C
BODIPY 50 mL 4-10°C
H33258 50 mL 4-10°C
Mounting agent 50 mL 4-10°C

<Protocol: 96-well plate format>

  1. Remove culture medium. Wash each well once with 100 uL of washing buffer.
  2. Add 50 uL of fixative solution to each well and fix overnight at room temperature.
  3. Remove fixative solution. Wash each well with 100 uL of washing buffer. (× 2 times)
  4. Add 50 uL of BODIPY to each well and incubate for 30 minutes at room temperature.
  5. Remove BODIPY. Add 50 uL of H33258 to each well and incubate for 30 minutes at room temperature.
  6. Remove H33258. Wash each well once with 100 uL of washing buffer.
  7. Add 50 uL of mounting agent to each well.
  8. Examine the sample by fluorescence microscope.

Note: For detection of lipid droplets, examine cells with an excitation filter of 498 nm and an emission filter of 503 nm.
For detection of nucleus, examine cells with an excitation filter of 352 nm and an emission filter of 461 nm.

<Application examples>
Visceral preadipocytes collected from a Spraguee Dawley rats (male) were differentiated into mature adipocytes and subjected to staining of lipid droplets (Figure 1) and nuclei (Figure 2).


Figure 1: BODIPY staining of lipid droplets

Figure 2: H33258 staining of nuclei

 

Product List

Product Name Cat# Quantity Price

Adipocyte Fluorescent Staining Kit

PMC-AK19F-COS 1SET

¥ 57,000
$ 760
€ 570

References
  • Sunao Takesita, Keisuke Kaji, Akira Kudo. Identification and Characterization of the New Osteoclast Progenitor with Macrophage Phenotypes Being Able to Differentiate into Mature Osteoclasts. JOURNAL OF BONE AND MINERAL RESEARCH Volume 15, Number 8 (2000). 1477-1488.

  • BEN A. A. SCHEVEN, JONE S. MILNE, SIMON P. ROBINS. A SEQUENTIAL CULTURE APPROACH TO STUDY OSTEOCLAST DIFFERENTIATION FROM NONADHERENT PORCINE BONE MARROW CELLS. In Vitro Cell. Dev. Biol. July-August (1998). Animal 34: 568-577

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