Description
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
Notice to Purchasers: This product is tobe used for Research Purposes Only. It is not to be used for Drug or Diagnostic Purposes, nor is it intended for Human Use. Primary cell products may not be resold, modified for resale, or used to manufacture commercial products without the express written consent of Primary Cell Co., Ltd.
Except as otherwise expressly set forth in this use manual, Primary Cell dose not make any representation or warranties or conditions of any kind, either express or implied, with respect to the products, orinformation disclosed hereunder, including, but not limited to, the implied warranties of merchantability, fit for a particular purpose, or noninfringement of the intellectual property rights of third parties.
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(*) 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
Notice to Purchasers: This product is tobe used for Research Purposes Only. It is not to be used for Drug or Diagnostic Purposes, nor is it intended for Human Use. Primary cell products may not be resold, modified for resale, or used to manufacture commercial products without the express written consent of Primary Cell Co., Ltd.
Except as otherwise expressly set forth in this use manual, Primary Cell dose not make any representation or warranties or conditions of any kind, either express or implied, with respect to the products, orinformation disclosed hereunder, including, but not limited to, the implied warranties of merchantability, fit for a particular purpose, or noninfringement of the intellectual property rights of third parties.
