Adipocyte Fluorescent Staining Kit

Assays/Cell Lineage Marker Assays/Adipocytes; Research Area/Development/Adipogenesis/Cell Lineage Marker Assays; Research Area/Cell Types/Adipocytes/Cell Lineage Marker Assays; Cells & Cell Culture/Assays/Cell Lineage Marker Assays/Adipocytes; Research Area/Cell Biology/Cell Metabolism/Cell Lineage Marker Assays; Assays/Adipocytes/Cell Lineage Marker Assays

Detection of Adipose Differentiation
The Adipocyte Fluorescent Staining Kit is designed to stain both lipid droplets and nuclei in adipocytes using BODIPY(R) (Invitrogen Corporation) and H33258, respectively. Furthermore, the amount of lipid per cell and cellshape can bequantified using imaging instrument such as IN Cell Analyzer 1000 (GE Healthcare Company)
(*) BODIPY is a registered trademark of Invitrogen Corporation.
Preparation of washing buffer: Add one Tablet for washing to 100 ml of distilled or deionized water and mix.
Note: This working solution can be stored at 4°C for 7 days.
Protocol (96-well plate format): 1. Remove culture medium. Wash each well once with 100uL of washing buffer.
2. Add 50uL of fixative solution to each well andfix overnight at room temperature.
3. Remove fixative solution. Wash each well with 100uL of washing buffer. (× 2 times)
4. Add 50uL of BODIPY to each well and incubate for 30 minutes at room temperature.
5. Remove BODIPY. Add 50uL of H33258to eachwell and incubate for 30 minutes at room temperature.
6. Remove H33258. Wash each well once with 100uL of washing buffer.
7. Add 50uL of mounting agent to each well.
8. Examine the sample by fluorescence microscope.
Note: For detection oflipid 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
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