{"product_id":"biolegend-422201","title":"Biolegend, 422201, Annexin V Binding Buffer, 100ml","description":"\u003cp\u003eAnnexin V Binding Buffer has been formulated for flow cytometric labeling of apoptotic cells with Annexin V reagents.  The buffer is provided as a ready-to-use solution.\u003cbr\u003e\n100ml\u003cbr\u003e\nStorage \u0026amp; Handling: Store between 2°C and 8°C.\u003cbr\u003e\nApplication: FC - Quality tested\u003cbr\u003e\nRecommended Usage: Annexin V Binding Buffer is recommended for use with Annexin V reagents. Cells should be resuspended in Annexin V Binding Buffer at a concentration 1x106\/ml and then combined with Annexin V reagents at the recommended volumes.\u003cbr\u003e\nApplication Notes: 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 \u0026amp; 420404), or Helix NP dyes (Cat. Nos. 425301, 425303, \u0026amp; 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.\u003cbr\u003e\nApplication References(PubMed link indicates BioLegend citation): Vermes I, et al. 1995. J. Immunol. Methods 184:39. Soon PS, et al. 2013. Endocr Relat Cancer. 20:1. PubMed Turner JE, et al. 2013. J Exp Med. 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PubMed\u003cbr\u003e\nGene ID: NA\u003cbr\u003e\nRegulatory Status: RUO\u003cbr\u003e\nOther Names: Annexin-V-Binding Buffer\u003cbr\u003e\nQ: How is your Annexin made and what sequence does it cover?\u003cbr\u003e\nA: It is made in E. coli, covering human aa Met1-Asp320.\u003cbr\u003e\nQ: How does pH and staining temperature affect Annexin V-Phosphatidylserine binding?\u003cbr\u003e\nA: Annexin-Phosphatidylserine binding is lost below pH 5.2 and with prolonged incubation over a temperature of 42°C.\u003cbr\u003e\nQ: Why do I need to use Annexin V Binding Buffer?\u003cbr\u003e\nA: Annexin V binding requires the presence of calcium in the solution.  So, we provide Annexin V Binding Buffer (cat # 422201), which is optimized for the best performance of Annexin V staining.\u003cbr\u003e\nQ: Can I use RPMI during Annexin V staining?\u003cbr\u003e\nA: It is best to follow protocol as described on the product data sheet. Moreover, RPMI 1640 has a relatively high concentration of phosphate and low calcium ion concentration, which negatively impacts Annexin binding to its target phosphatidylserine (PS). Measurement of cell death by using Annexin V may also be significantly affected by time of incubation on ice, calcium concentration, and type of medium.\u003cbr\u003e\nQ: Can I freeze Annexin V conjugates?\u003cbr\u003e\nA: It should not be frozen as it will lead to loss of biological activity due to dimerization.\u003cbr\u003e\nQ: Is Annexin V suitable for conjugation with the Maxpar® kit for CyTOF®?\u003cbr\u003e\nA: Maxpar® Labeling kits require the protein to be partially reduced, so the metal chelate can be introduced through an SH group in the hinge region of the reduced antibody. Human Annexin V contains only one Cysteine which was reported to be chemically inactive. Thus, the Maxpar® labeling protocol would not work with Annexin V, unless a free –SH group can be introduced to Annexin V.  For more information regarding SH-mediated conjugation of Annexin V please consult published papers such as this one.\u003c\/p\u003e","brand":"Biolegend","offers":[{"title":"Default Title","offer_id":46864299032745,"sku":"422201","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/es\/products\/biolegend-422201","provider":"Iright","version":"1.0","type":"link"}