Antibody data
- Antibody Data
- Antigen structure
- References [3]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [4]
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- Product number
- 63-1178-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD117 (c-Kit) Monoclonal Antibody (104D2), Super Bright™ 600, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- The 104D2 monoclonal antibody reacts with human CD117, also known as c-Kit, Steel factor receptor, and stem cell factor receptor. A member of the tyrosine kinase receptor family, this 145 kDa molecule is expressed by hematopoietic progenitor cell subsets and mast cells. The interaction of c-Kit and Steel factor promotes proliferation and differentiation of hematopoietic progenitor cells and mast cell differentiation. CD117 is also expressed by melanocytes and plays a role in signaling and activation of these cells. This 104D2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.25 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 2-8°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorochrome performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 104D2
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Chemically-Defined, Xeno-Free, Scalable Production of hPSC-Derived Definitive Endoderm Aggregates with Multi-Lineage Differentiation Potential.
IL1RAP potentiates multiple oncogenic signaling pathways in AML.
PPAR-α and glucocorticoid receptor synergize to promote erythroid progenitor self-renewal.
Sahabian A, Sgodda M, Naujok O, Dettmer R, Dahlmann J, Manstein F, Cantz T, Zweigerdt R, Martin U, Olmer R
Cells 2019 Dec 4;8(12)
Cells 2019 Dec 4;8(12)
IL1RAP potentiates multiple oncogenic signaling pathways in AML.
Mitchell K, Barreyro L, Todorova TI, Taylor SJ, Antony-Debré I, Narayanagari SR, Carvajal LA, Leite J, Piperdi Z, Pendurti G, Mantzaris I, Paietta E, Verma A, Gritsman K, Steidl U
The Journal of experimental medicine 2018 Jun 4;215(6):1709-1727
The Journal of experimental medicine 2018 Jun 4;215(6):1709-1727
PPAR-α and glucocorticoid receptor synergize to promote erythroid progenitor self-renewal.
Lee HY, Gao X, Barrasa MI, Li H, Elmes RR, Peters LL, Lodish HF
Nature 2015 Jun 25;522(7557):474-7
Nature 2015 Jun 25;522(7557):474-7
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of normal human peripheral blood cells with the Human Hematopoietic Lineage eFluor® 450 Cocktail (Product # 22-7775-72), Anti-Human CD34 FITC (Product # 11-0349-42) and Mouse IgG1 K Isotype Control Super Bright 600 (Product # 63-4714-82) (left) or Anti-Human CD117 (c-Kit) Super Bright 600 (right). Lineage negative/low cells were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 1 Generation and characterization of DE aggregates in Erlenmeyer flasks. ( a ) Schematic overview of definitive endoderm differentiation in suspension. ( b ) Cell density analyzed at day 0 and day 3 of differentiation (n = 4-6). ( c ) Representative brightfield image of iPSC aggregates at day 0 and day 3 of differentiation (n = 4-6). ( d ) Aggregate diameters measured at day 0 and day 3 of differentiation. ( e ) Definitive endoderm efficiency quantification based on flow cytometry analysis of CXCR4/c-Kit and CXCR4/EpCAM double positive cells at day 3 of differentiation (n = 12-15). ( f ) qRT-PCR analysis of FOXA2 and SOX17 expression at day 3 of differentiation (n = 6-9). ( g ) Immunostaining of SOX17 (white) and FOXA2 (red), and nuclear stain DAPI (blue) of DE aggregates at day 3 of differentiation (n = 3). Scale bar, 100 mum. Each value of gene expression was first normalized to the reference genes and then to day 0 undifferentiated cells. Values are represented as the mean +- SEM. All n values correspond to independent experiments.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Extended Figure 7 a, Real-time RT-PCR analysis of Kit gene expression in wild-type and PPARalpha -/- mouse BFU-E cells untreated or treated with DEX with or without addition of GW7647 (* p < 0.05; Student t test. Error bars represent mean +- S.D. from three independent experiments.); b , Human CD34+ cells were treated with or without GW7647 as described in the legend to Figure 2 . (top) At day 9 of culture, cell surface KIT and CD71 expression were analyzed by flow cytometry. (bottom) A representative histogram of KIT expression in cells treated or untreated with GW7647; c , ChIP-Seq occupancy signal map of GR and PPARalpha across the Kit locus in BFU-E cells.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Extended Figure 2 Human CD34 + Erythroid Differentiation System a , Total CFU-E colonies formed during day 0-9. CFU-E colony numbers were quantified by plating 1000 cells from various time points during day 0-9 of the human CD34 + erythroid culture on methylcellulose. CFU-E colonies were quantified after 12-14 days. Total CFU-E colony numbers in culture under conditions without GW7647 (black line) or with GW7647 (red line) were calculated using the total cell numbers at corresponding time points in Figure 2a. b , Human CD34+ cells were treated at day 1 with 100 nM GW7647 with or without DEX at the concentration indicated in the figure. At day 6, total cell numbers were counted and cells were collected for BFU-E colony assays. c , Protein expression of PPARalpha demonstrating shRNA knock-down efficiency via lentiviral transduction. LacZ shRNA is used as a control. shRNA-1 and -2 are both specific for PPARalpha . shRNA-2 has higher knock-down efficiency. d , Cell pellets of 1 million cells demonstrating hemoglobin accumulation during the differentiation process. e , Flow cytometry analyses of erythroid markers during the 21-day human CD34 + erythroid culture. (top row) c-kit vs. CD235a; (middle row) CD71 vs. CD235a. Note the sequential induction of c-kit, CD71 and CD235a, as well as the sequential down-regulation of c-kit and CD71, (bottom row) Enucleated reticulocytes are CD235a + Hoechst - , nuclei are CD235a - Hoechst + , and nucleated erythroblasts are CD235a + Hoechst + .
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4. IL1RAP physically interacts with cytokine receptors FLT3 and c-KIT in human AML cells. (A) Coimmunoprecipitation of HA-tagged IL1RAP in protein lysates from 293T cells transfected with IL1RAP-HA and FLT3-myc fusion vectors. Immunoblot (IB) was performed for IL1RAP and FLT3. (B) Coimmunoprecipitation of endogenous FLT3 in protein lysates from THP-1 cells. Immunoblot was performed for FLT3 and IL1RAP. (C) Fluorescence energy transfer (FRET) between IL1RAP and FLT3. THP-1 (left) or MOLM-13 (right) cells were stimulated with 200 ng/ml rhIL-1beta or 100 ng/ml rhFLT3L. For C and G, cells were stained with fluorescent antibodies against the indicated proteins for flow cytometry analysis and FRET was determined for cells expressing both receptors. Data represent the mean +- SD of three independent experiments. (D) Coimmunoprecipitation of HA-tagged IL1RAP in protein lysates from 293T cells transfected with IL1RAP-HA and FLT3-ITD-myc fusion vectors. Immunoblot was performed for IL1RAP and FLT3. (E) Coimmunoprecipitation of HA-tagged IL1RAP in protein lysates from 293T cells transfected with IL1RAP-HA and c-KIT-myc fusion vectors. Immunoblot was performed for IL1RAP and c-KIT. (F) Coimmunoprecipitation of endogenous c-KIT in protein lysates from THP-1 cells. Immunoblot was performed for c-KIT and IL1RAP. (G) FRET between IL1RAP and c-KIT. THP-1 (left) or HEL (right) cells were stimulated with 200 ng/ml IL-1beta or 100 ng/ml SCF. Data represent the mean +- SD of t