Luminex

Location:Home - Technical - Protocol - Luminex

Proper sample handling for Luminex

Cell culture supernatant

1. Stimulate cells as desired in the appropriate cell culture flasks. Cells should be in log phase growth when harvesting supernatant.

2. Using sterile technique, remove the desired volume of conditioned cell culture medium with a pipette and transfer the medium to clean polypropylene microcentrifuge tubes.

3. Centrifuge the medium at 14,000 rpm for 10 minutes at 4°C in a refrigerated microcentrifuge to remove any cells or cellular debris.

4. Aliquot the clarified medium into clean polypropylene microcentrifuge tubes. 5. If supernatant is to be analyzed at a later date, dispense into aliquots and store at –80°C.

 

Serum

1. Collect blood samples in pyrogen- and endotoxin-free tubes.

2. Allow whole blood to sit at room temperature for 15–30 minutes to clot.

3. Spin at 1,000–2,000 x g for 10 minutes at 4°C in a refrigerated centrifuge to separate the cells.

4. Transfer the supernatant to a clean, chilled polypropylene tube with a sterile Pasteur pipette. 5. If serum is to be analyzed at a later date, dispense into aliquots in polypropylene microcentrifuge tubes and store at –80°C. When possible, avoid the use of hemolyzed or lipemic serum.

 

Plasma

1. Remove cells from plasma samples by centrifugation at 2,000 x g for 10 minutes at 4°C in a refrigerated centrifuge. Centrifugation at this force is necessary to deplete platelets from the sample.

2. Transfer the supernatant to a clean, chilled polypropylene tube with a sterile Pasteur pipette.

3. If the plasma is to be analyzed at a later date, dispense into aliquots in polypropylene microcentrifuge tubes and store at –80°C.

 

Tissue homogenate

1. Add protease inhibitors to Tissue Extraction Reagent I just before use.

2. Weigh tissue sample.

3. Add 10 mL of Tissue Extraction Reagent I per 1 gram of tissue.

4. Homogenize the tissue (best practice is to use ice-cold buffers and homogenize tissue on ice). 5. Centrifuge the sample at 10,000 rpm for 5 minutes at 4°C to pellet the tissue debris. 6. Collect the supernatant. The sample is ready to be used.* 7. If the sample is to be stored, dispense into aliquots and freeze at –80°C.

Share:
Back
2020无码最新国产在线观看-无码国模国产在线观看-最新国模无码国产在线视频_福利