Biolegend, 285490, Spark UV™ 387 anti-human CD95 (Fas) (Flexi-Fluor™) Antibody, 50μg

CD95 is a 45 kD single chain type I glycoprotein also known as Fas, APO-1, and TNFRSF6. It is a member of the TNF receptor superfamily. CD95 is expressed on T and B lymphocytes, monocytes, neutrophils, and fibroblasts. CD95 expression is upregulated by activation. The extracellular region of CD95 binds to CD178 (Fas ligand). CD178 binding to CD95 induces apoptosis and has been shown to play a role in the maintenance of peripheral tolerance.
50μg
Verified Reactivity: Human, Cynomolgus, Rhesus
Reported Reactivity: African Green, Baboon, Capuchin Monkey, Chimpanzee, Common Marmoset, Cotton-topped Tamarin, Pigtailed Macaque, Sooty Mangabey
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: CD95 transfected L cells
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark UV™ 387 under optimal conditions.
Concentration: 0.2 mg/mL
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
Recommended Usage: Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL. For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. * Spark UV™ 387 has a maximum excitation of 351 nm and a maximum emission of 387 nm.
Excitation Laser: Ultraviolet Laser (355 nm)
Application Notes: The DX2 antibody is useful for inducing apoptosis of Fas-positive cells. Additional reported applications (for the relevant formats) include: in vitro induction of apoptosis3 (DX2 antibody is required to be cross-linked for effective induction of apoptosis) and immunohistochemical staining4,5 of acetone-fixed frozen tissue sections and formalin-fixed paraffin-embedded tissue sections. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 305655 and 305656). Note: EOS9.1 antibody can induce apoptosis without cross-linking.
Additional Product Notes: For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.
Application References(PubMed link indicates BioLegend citation): Schlossman S, et al. Eds.1995. Leucocyte Typing V. Oxford University Press. New York. Kishimoto T, et al. Eds. 1997. Leucocyte Typing VI. Garland Publishing Inc. New York. Cifone M, et al. 1994. J. Exp. Med. 180:1547. (Apop) Zietz C, et al. 2001. Am. J. Pathol. 159:963. (IHC) Sergi C, et al. 2000. Am. J. Pathol. 156:1589. (IHC) Xie S, et al. 2010. J. Immunol. 184:2289. (FC) PubMed Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Sestak K, et al. 2007. Vet. Immunol. Immunopathol. 119:21. Rout N, et al. 2010. PLoS One 5:e9787. (FC) Dixit N, et al. 2012. J. Immunol. 189:5954. PubMed
RRID: AB_3716997 (BioLegend Cat. No. 285490)
Structure: TNFR superfamily, type I transmembrane protein, 45 kD
Distribution: T cells and B cells, monocytes, neutrophils, fibroblasts, expression level upregulated on activated lymphocytes
Function: Mediates apoptosis
Ligand/Receptor: Fas ligand (CD178)
Cell Type: B cells, Fibroblasts, Lymphocytes, Monocytes, Neutrophils, T cells
Biology Area: Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Immunology, Neuroscience
Molecular Family: CD Molecules
Antigen References: 1. Krammer P, et al. 1994. Immunol. Rev. 142:175. 2. Nagata S, et al. 1995. Science 267:1449.
Gene ID: 355
UniProt: View information about CD95 on UniProt.org
Clone: DX2
Regulatory Status: RUO
Workshop: VI C-64
Other Names: Fas, APO-1, TNFRSF6
Isotype: Mouse IgG1, κ
Q: What are Flexi-Fluors?
A: Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer.
Q: How quickly will I receive my order?
A: We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.
Q: How are Flexi-Fluors different from regular catalog products?
A: Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.
Q: How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?
A: Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
Q: I can’t find the antibody-dye combination that I need. When will it be available?
A: We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services.
Q: I need help to validate the performance of my Flexi-Fluor. Who should I contact?
A: Please get in touch with Technical Service for assistance.
Q: Can I order more than 50 μg of a Flexi-Fluor?
A: Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.
Q: What is the expiration date of my Flexi-Fluor?
A: Expiration dates can be found on the vial label or by using our CoA lookup tool.
Q: What if my requested antibody-dye combination doesn't perform as expected?
A: Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.