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Flow cytometric analysis of Axl expression on human HeLa cells. Cells from the human HeLa (Cervical adenocarcinoma, ATCC CCL-2) cell line were stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD OptiBuild™ BV421 Mouse Anti-Human Axl (Cat. No. 747864, solid line histogram) at 0.25 µg/test. The fluorescence histogram showing Axl expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable HeLa cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. The above is qualification data only and does not represent a specific OptiBuild™ lot.
BD OptiBuild™ BV421 Mouse Anti-Human Axl
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
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The 108724 monoclonal antibody specifically recognizes Axl which is also known as adhesion related kinase (ARK), JTK11, Tyro7, or UFO. Tyro-3, Axl, and Mer constitute the TAM subfamily of receptor tyrosine kinases (RTK). Axl is a single-pass type I transmembrane glycoprotein comprised of an extracellular region with two immunoglobulin (Ig)-like domains and two fibronectin type III (FNIII) domains, a transmembrane segment and a conserved intracellular tyrosine kinase domain. Axl is variably expressed by multiple cell types including monocytes, macrophages, dendritic cells, NK cells, platelets, endothelial cells, vascular smooth muscle cells, and fibroblasts. Axl binds to the vitamin K-dependent Growth arrest-specific protein 6 (Gas6) through its extracellular Ig-like domains. Ligand binding leads to receptor dimerization, and autophosphorylation of tyrosine residues within the cytoplasmic Axl domains. This results in the activation of downstream signaling pathways that control cellular adhesion, aggregation, phagocytosis/efferocytosis, proliferation, survival, and migration. Through these activities, Axl plays major roles in development, the regulation of hematopoiesis and immunity, and ensuring the integrity of the vascular system. Abnormal expression of Axl has been observed in various cancers and myeloproliferative disorders and by tumor cell lines.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
Development References (8)
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Gould WR, Baxi SM, Schroeder R, et al. Gas6 receptors Axl, Sky and Mer enhance platelet activation and regulate thrombotic responses.. J Thromb Haemost. 2005; 3(4):733-41. (Biology). View Reference
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Janssen JW, Schulz AS, Steenvoorden AC, et al. A novel putative tyrosine kinase receptor with oncogenic potential.. Oncogene. 1991; 6(11):2113-20. (Biology). View Reference
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Lemke G, Rothlin CV. Immunobiology of the TAM receptors. Nat Rev Immunol. 2008; 8(5):327-336. (Biology). View Reference
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Linger RM, Keating AK, Earp HS, Graham DK. TAM receptor tyrosine kinases: biologic functions, signaling, and potential therapeutic targeting in human cancer.. Adv Cancer Res. 2008; 100:35-83. (Biology). View Reference
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Rothlin CV, Carrera-Silva EA, Bosurgi L, Ghosh S. TAM receptor signaling in immune homeostasis.. Annu Rev Immunol. 2015; 33:355-91. (Biology). View Reference
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Shiozawa Y, Pedersen EA, Patel LR, et al. GAS6/AXL axis regulates prostate cancer invasion, proliferation, and survival in the bone marrow niche. Neoplasia. 2010; 12(2):116-127. (Clone-specific: Flow cytometry). View Reference
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Taichman RS, Patel LR, Bedenis R, et al. GAS6 receptor status is associated with dormancy and bone metastatic tumor formation.. PLoS ONE. 2013; 8(4):e61873. (Clone-specific: Flow cytometry). View Reference
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Yumoto K, Eber MR, Wang J, et al. Axl is required for TGF-β2-induced dormancy of prostate cancer cells in the bone marrow.. Sci Rep. 2016; 6:36520. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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