12-1178-41

antibody from Invitrogen Antibodies

Targeting: KIT C-Kit, CD117, PBT, SCFR

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

12-1178-41

Provider

Invitrogen Antibodies

Product name

CD117 (c-Kit) Monoclonal Antibody (104D2), PE, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

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.

Conjugate

Yellow dye

Antibody clone number

104D2

Concentration

5 µL/Test

References

Submitted references

Chemically-Defined, Xeno-Free, Scalable Production of hPSC-Derived Definitive Endoderm Aggregates with Multi-Lineage Differentiation Potential.

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)

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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

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Induced pluripotent stem cell-based mapping of β-globin expression throughout human erythropoietic development.

Vanuytsel K, Matte T, Leung A, Naing ZH, Morrison T, Chui DHK, Steinberg MH, Murphy GJ
Blood advances 2018 Aug 14;2(15):1998-2011

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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

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of normal human peripheral blood cells with Anti-Human CD34 FITC (Product # 11-0349-42) and Mouse IgG1 K Isotype Control PE (Product # 12-4714-81) (left) or Anti-Human CD117 (c-Kit) PE (right). CD45 dim cells in the lymphocyte gate were used for analysis.

Conjugate

Yellow dye

Other assay

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.

Conjugate

Yellow dye

Supportive validation

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.

Conjugate

Yellow dye

Supportive validation

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 + .

Conjugate

Yellow dye