Biolegend, 640953, APC/Fire™ 750 Annexin V
100tests
Catalog Number: 640953
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| Verified Reactivity | Human, Mouse, Rat |
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| Reported Reactivity | Other Species |
| Formulation | Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA) |
| Preparation | The antibody was purified by affinity chromatography and conjugated with APC/Fire™ 750 under optimal conditions. |
| Concentration | Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.) |
| Storage & Handling | The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze. |
| Application | FC - Quality tested |
| Recommended Usage | Each lot of this product is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric application, the suggested use of this reagent is 5 - 15 µg per 100,000 - million cells in a 100 µL volume of Annexin V Binding Buffer (Cat No. 422201). It is recommended that the reagent be titrated for optimal performance for each application. * APC/Fire™ 750 has a maximum excitation of 650 nm and a maximum emission of 774 nm. |
| Excitation Laser | Red Laser (633 nm) |
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| Application Notes |
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| Annexin V Staining Wash cells twice with cold BioLegend Cell Staining Buffer (Cat. No. 420201) and then resuspend cells in Annexin V Binding Buffer (Cat. No. 422201) at a concentration of 1x106 cells/mL. Transfer 100 µL of cell suspension in 5 mL test tube. Add 5 µL of fluorochrome conjugated Annexin V. Stain with a viability dye, such as PI (Cat. No. 421301), 7-AAD (Cat. Nos. 420403 & 420404), or Helix NP dyes (Cat. Nos. 425301, 425303, & 425305), if desired. Gently vortex the cells and incubate for 15 min at RT (25°C) in the dark. Add 400* µL of Annexin V Binding Buffer (Cat. No. 422201) to each tube. *For more concentrated samples, add a minimum of 200 µl of Annexin V Binding Buffer in this step. Analyze by flow cytometry. For a better experience detecting apoptosis, we now recommend Apotracker™. Cell staining with Apotracker™ is Calcium independent. Thus, no special buffers are required, and the protocol can be shortened for single-step co-staining with other reagents. |
| Application References(PubMed link indicates BioLegend citation) | Koopman G, et al. 1994. Blood 84:1415. Vermes I, et al. 1995. J. Immunol. Methods 184:39. Dachary-Prigent J, et al. 1993. Blood 81:2554. Sekine C, et al. 2009. Int Immunol. PubMed Grujic M, et al. 2010. J. Immunol. 185:1730. PubMed |
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| Product Citations | de Winde CM, et al. 2021. iScience. 24:102976. PubMed Parayath NN, et al. 2020. Nat Commun. 4.680555556. PubMed |
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| RRID | not an antibody (BioLegend Cat. No. 640952) not an antibody (BioLegend Cat. No. 640953) |
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| Biology Area | Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Neuroscience |
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| Gene ID | 308 |
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| UniProt | View information about Annexin V on UniProt.org |
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| Regulatory Status | RUO |
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| Other Names | Annexin A5, Dead, Apoptosis |
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