Biolegend, 285540, Spark YG™ 581 anti-human CD117 (c-kit) (Flexi-Fluor™) Antibody, 50μg

CD117 is a 145 kD protein tyrosine kinase also known as c-Kit. It is a receptor for stem cell factor or c-Kit ligand. CD117 is expressed on pluripotent hematopoietic progenitor cells (approximately 1-4% bone marrow cells), mast cells, and acute myeloid leukemia cells (AML). CD117 binding of c-Kit ligand induces phosphorylation of CD117 and stimulates proliferation and survival of primitive hematopoietic stem cells as well as erythroid-committed and granulo-monocytic committed cells.
50μg
Verified Reactivity: Human
Reported Reactivity: Cynomolgus, Cow
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: MOLM-1 megakaryocytic cell line
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark YG™ 581 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 YG™ 581 has a maximum excitation of 562 nm and a maximum emission of 581 nm.
Excitation Laser: Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes: The 104D2 antibody does not block binding of c-Kit ligand. Additional reported applications (for the relevant formats) include: immunoprecipitation1, immunofluorescence microscopy1, and spatial biology (IBEX)4,5.
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): Broudy VC, et al. 1999. Blood 94:1979. (IF, IP) Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Nagano M, et al. 2007. Blood 110:151. (FC) PubMed Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID: AB_3720651 (BioLegend Cat. No. 285540)
Structure: Growth factor receptor with tyrosine kinase activity, subclass III, approximately 145 kD
Distribution: Pluripotent hematopoietic progenitor cells (approximately 1-4% bone marrow cells), mast cells, acute myeloid leukemic cells (AML)
Function: Growth factor receptor for stem cell factor. Induces proliferation and survival of primitive hematopoietic progenitors. Potent inducer of proliferation in erythroid-committed progenitor cells. Defects in CD117 have been linked to severe anemia and a decreased number of hematopoietic progenitor cells.
Ligand/Receptor: c-Kit ligand
Modification: Multiple phosphorylation sites
Cell Type: Embryonic Stem Cells, Hematopoietic stem and progenitors, Leukemia, Mast cells, Mesenchymal Stem Cells
Biology Area: Immunology, Stem Cells
Molecular Family: CD Molecules
Antigen References: 1. Giebel LB, et al. 1992. Oncogene 7:2207. 2. Furitsu T, et al. 1993. J. Clin. Invest. 92:1736.
Gene ID: 3815
UniProt: View information about CD117 on UniProt.org
Clone: 104D2
Regulatory Status: RUO
Other Names: Stem cell factor receptor, c-kit, mast cell growth factor receptor, steel factor receptor
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.