Biolegend, 391507, Human GPI-APD Cocktail for WBC with Control Cells
50tests
Catalog Number: 391507
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Category
Pack Size2 vials
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| Verified Reactivity | Human |
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| Concentration | Veri-Cells™ Leukocytes ≥ 1x106 cells per vial (lyophilized) GPI-APD cocktail for WBC: lot-specific |
| Storage & Handling | Store the kit at 2°C - 8°C upon receipt. Do not open sealed pouch until ready to use. Unstained Veri-Cells™ Leukocytes reconstituted in Buffer A Plus or GPI Buffer can be stored separately up to five days at 2°C - 8°C. Do not mix the two reconstituted Veri-Cells™ Leukocytes until ready to use as Veri-Cells™ Control Cells. |
| Application | FC - Quality tested |
| Recommended Usage | Veri-Cells™ Leukocytes: 50 μL per test (after reconstitution with 325 μL of Veri-Cells™ Buffer A Plus or GPI Buffer). Once reconstituted, let the cells sit for at least 30 minutes at room temperature prior to staining. GPI-APD cocktail for WBC: the suggested use of this reagent is 20 μL per 50 μL reconstituted Veri-Cells™ or 100 μL of whole blood for flow cytometric staining. It is recommended that the reagent be titrated for optimal performance for each application. Note: Veri-Cells™ Buffer A plus and GPI Buffer may show precipitation over time; however, this is normal and does not affect the performance of the buffer. |
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| Application Notes |
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| Veri-Cells™ Control Cell Protocol Remove the kit from the fridge and allow it to warm to room temperature for 5 minutes. Keep unused kit components stored in the fridge. Reconstitute 1 Veri-Cells™ Leukocytes vial with 325 µL of Buffer A Plus. Reconstitute a second Veri-Cells™ Leukocytes vial with 325 µL of GPI Buffer. Label each vial with the reconstitution buffer (“Buffer A Plus” and “GPI Buffer”) Incubate the 2 vials at room temperature for 30 minutes. Prepare two 5 mL FACS tubes (not provided). Label one “Buffer A Plus” and the other “GPI Buffer.” Add 20 μL of the GPI-APD Cocktail for WBC to each tube Add 50 μL of Veri-Cells™ Leukocytes reconstituted in Buffer A Plus or GPI Buffer to the appropriately labeled tube. Incubate the cells at room temperature and in the dark for about 15 minutes. Add 2 mL of Cell Staining Buffer (Cat. No. 420201). Centrifuge at 350 x g for five minutes. Discard supernatant. Wash one more time with 2 mL of Cell Staining Buffer. Resuspend cell pellet in 0.2 - 0.4 mL of Cell Staining Buffer. Combine the two tubes to make one tube of “Veri-Cells™ Control Cells.” Acquire cells immediately (along with samples) on a flow cytometer or store at 4°C, in the dark, for no longer than 12 hours. Note: Unstained Veri-Cells™ Leukocytes reconstituted in Buffer A Plus or GPI Buffer can be stored separately up to five days at 2°C - 8°C. Whole Blood Cell (WBC) Protocol Prepare 1X RBC Lysis/Fixation solution by diluting 1 part of RBC Lysis/Fixation solution with 9 parts of room temperature deionized water. Prepare single color controls (listed in the Recommended Reagents) using either WBC or Compensation Beads (Cat. No. 424602). Mix 100 μL of whole blood with 20 μL of the GPI-APD Cocktail for WBC. Incubate at room temperature, in the dark, for 15 minutes. Add 2.0 mL of 1X RBC Lysis/Fixation solution, gently mixing each tube immediately. Incubate at room temperature, in the dark, for 15 minutes. Centrifuge at 350 x g for five minutes. Discard supernatant. Add 2 mL of Cell Staining Buffer. Centrifuge at 350 x g for five minutes. Discard supernatant. Wash one more time with 2 mL of Cell Staining Buffer. Resuspend cell pellet in 0.3 - 0.5 mL of Cell Staining Buffer. Acquire cells immediately (along with Veri-Cells™ Control Cells) on a flow cytometer or store at 4°C, in the dark, for no longer than 12 hours. |
| Cell Type | Granulocytes, Monocytes |
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| Molecular Family | CD Molecules |
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| Gene ID | NA |
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| Regulatory Status | RUO |
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| Other Names | Glycosylphosphatidylinositol (GPI)-anchored protein, control cells, Veri-Cells control |
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