Biolegend, 423303, Cell Activation Cocktail (with Brefeldin A)
100microl
Catalog Number: 423303
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Pack Size4 vials
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| Formulation | Cell Activation Cocktail (with Brefeldin A) is composed of PMA, ionomycin, and Brefeldin A in DMSO (500X). |
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| Concentration | Each vial of this cocktail contains phorbol-12-myristate 13-acetate (40.5 µM), ionomycin (669.3 µM), and Brefeldin A (2.5 mg/ml) in DMSO (500X). |
| Storage & Handling | Store cocktail at -70°C upon receipt. Protect from light. Avoid repeat freeze/thaw cycles. |
| Application | Activ, FC, ICFC - Quality tested |
| Recommended Usage | Cat. No. 423303 contains 100 µL of cocktail, formulated at 500X, that can be used for activating 50 mL of cells of various types. Cat. No. 423304 contains 400 µL (4 vials x 100 µL per vial) of cocktail, formulated at 500X, that can be used for activating 200 mL of cells of various types. |
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| Application Notes |
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| Cell Activation Protocol: 1. Resuspend cells of interest in desired cell culture medium. Recommended cell suspension is between 1-2 x 106 cells/mL, although effective activation can be obtained with higher or lower concentration. 2. Completely thaw vials in 37°C water bath. 3. Add 2 µL of the cocktail to each mL of cell suspension. 4. Incubate cells at 37°C in a CO2 incubator for 6 hours or the time period of interest. 5. Harvest activated cells and centrifuge at 350g for 5 minutes. Discard supernatant. 6. Add 2.5 mL of Cell Staining Buffer (Cat. No. 420201) to the cell pellet, mix by vortex or pipetting, then centrifuge at 350g for 5 minutes. Discard supernatant. 7. Repeat step 6. 8. Cells are now ready to be tested for surface markers or proteins of interest. Note: Activation of cells overnight (>12 hours) with Brefeldin A is generally not recommended due to some level of toxicity. Under these conditions, it is recommended to use Cell Activation Cocktail without Brefeldin A (Cat. No. 423301), and then add monensin (Cat. No. 420701) at appropriate concentrations. This product is temperature sensitive; repeated freeze/thaw is not recommended. Intracellular and surface flow staining, as well as ELISA, can be performed following activation of cells with this cocktail. |
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| Gene ID | NA |
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| Regulatory Status | RUO |
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| Other Names | Cell Stimulation Cocktail, PMA/Ionomycin |
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