Biolegend, 313203, PE anti-human CD117 (c-kit) Antibody, 25tests

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.
25tests
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 and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography, and conjugated with PE 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 CD117 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 SB - Reported in the literature, not verified in house
Recommended Usage: Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.
Excitation Laser: Blue Laser (488 nm)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: Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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
Product Citations: Matkovic Leko I, et al. 2023. Nat Protoc. . PubMed Sun J, et al. 2008. J Biol Chem. 283:27444. PubMed Sun J, et al. 2018. Oncogenesis. 7:48. PubMed Zhao Y, et al. 2016. Cell Rep. 16:2003-2016. PubMed Sun J, et al. 2009. J Biol Chem. 284:11039. PubMed Niwa A, et al. 2011. PLoS One. 6:e22261. PubMed Cho K, et al. 2017. Biochem Biophys Res Commun. 10.1016/j.bbrc.2017.04.141. PubMed Cruse G, et al. 2015. Mol Biol Cell. 26:1711. PubMed Taguchi A, et al. 2017. Cell Stem Cell. 21:730. PubMed Lecluze E, et al. 2020. Hum Reprod. 35:1099. PubMed Shen C, et al. 2021. Front Immunol. 12:680055. PubMed Zhang J, et al. 2019. Dev Cell. 48:329. PubMed Suzuki S, et al. 2021. STAR Protocols. 2:100683. PubMed Schielke L, et al. 2022. Front Immunol. 13:916701. PubMed Fang F, et al. 2021. Cell Rep. 37:109981. PubMed Zeng Z, et al. 2021. Nat Commun. 12:3641. PubMed Smith-Díaz CC, et al. 2021. Front Oncol. 11:709543. PubMed López-Sanz C, et al. 2022. STAR Protoc. 3:101755. PubMed Bender R, et al. 2016. PLoS Pathog. 12: 1005641. PubMed Zhang Y et al. 2018. Cell stem cell. 23(4):516-529 . PubMed Pan C, et al. 2019. Int J Mol Med. 44:1629. PubMed Mpakou V, et al. 2021. Exp Ther Med. 1.010416667. PubMed Samuel RM, et al. 2020. Cell Stem Cell. 27:876. PubMed Bennstein SB, et al. 2020. eLife. 9:e55232.. PubMed Wang G, et al. 2018. Int J Mol Med. 41:791. PubMed
RRID: AB_314982 (BioLegend Cat. No. 313203) AB_314983 (BioLegend Cat. No. 313204)
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 type of PE do you use in your conjugates?
A: We use R-PE in our conjugates.
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Q: Are other fluorophores compatible with IBEX?
A: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
Q: The same antibody works in one tissue type but not another. What is happening?
A: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.