Pancreatic Beta-cell Staining Kit - Cosmo Bio Co.,Ltd.

Antibodies

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Pancreatic Beta-cell Staining Kit

Catalog No.: PMC-AK11F-COS
Size: 1SET
Price: ¥57000
$760
catalog info: Catalog 2012-p143
Storage: 4C DNF
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Purpose: Pancreatic -cell Fluorescent Staining Kit (PMC-AK11F-COS)@is designed to stain insulin-producing -cells, using an anti-insulin antibody. This product is available for mouse and rat -cells.using anti-insulin antibody. This product is available for mouse and rat -cells.
component: Washing Buffer (100 mL) x 1 bottle
Blocking Buffer (2 mL) x 1 bottle
Anti-insulin Antibody (20 uL) x 1 vial
Fluorescein Secondary Antibodies (20 uL) x 1 vial
One kit contains regents for 10 samples
Species Cross-Reactivity: rat, mouse
Additional Materials Required
Efixative solution 4% Paraformaldehyde Phosphate Buffer Solution

Other:

 Protocol 
1. Transfer pancreatic cells (1.0~105 cells and above) or 10-20 pancreatic islets into a suitable centrifuge tube such as 1.5 ml centrifuge tube.
2. Centrifuge at 2,000 x g for 3 minutes and remove the supernatant.
3. Fix cellswith 0.5 ml of fixative solution at 4C over night.
4. Centrifuge at 10,000 x g for 3 minutes and remove the fixative solution.
5. Add 1 ml of Washing Buffer to centrifuge tube and centrifuge at 10,000 x g for 3 minutes, and then remove the WashingBuffer. (x 3 times)
6. After removing the Washing Buffer, add 100 ul of Blocking Buffer to the centrifuge tube and leave for 30 minutes at room temperature.
7. Centrifuge at 10,000 x g for 3 minutes and remove the Blocking Buffer.
8. Add100ul of Blocking Buffer to the centrifuge tube and add 2 ul of Anti-insulin antibody to the tube. Mix gently using a micropipette and leave the tube at room temperature for one hour.
9. Centrifuge at 10,000 x g for 3 minutes and remove the Anti-insulin antibody.
10. Add 1 ml of Washing Buffer to the centrifuge tube and centrifuge at 10,000 x g for 3 minutes. Remove the Washing Buffer (x 3 times).
11. After removing the Washing Buffer, add 100 ul of Blocking Buffer to centrifuge tube and add 2 ul ofFluorescein Secondary Antibodies to the tube. Mix gently using a micropipette and leave the tube at room temperature for one hour.
12. Centrifuge at 10,000 x g for 3 minutes and remove the Anti-insulin antibody.
13. Add 1 ml of WashingBuffer to the centrifuge tube and centrifuge at 10,000 x g for 3 minutes, and remove the Washing Buffer. (x 3 times)
14. Examine cells with an excitation filter of 459 nm and an emission filter of 520 nm .



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