C kit apc

Manufactured by BD

Sourced in United States

C-Kit-APC is a flow cytometry reagent that detects c-kit (CD117) expression on the surface of cells. C-kit is a receptor tyrosine kinase that is important for the development and function of various cell types, including hematopoietic stem cells, mast cells, and germ cells. The APC conjugate allows for the fluorescent detection of c-kit-positive cells using flow cytometry.

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Lab products found in correlation

Pe cy7 sca, by BD (4 mentions) Apc cy7 cd48, by Thermo Fisher Scientific (4 mentions) Facsaria cell sorter, by BD (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Lineage cell depletion kit, by Miltenyi Biotec (3 mentions) Fetal bovine serum (fbs), by Merck Group (3 mentions) Hepes, by Thermo Fisher Scientific (3 mentions) Pacific blue cd150, by BioLegend (3 mentions) B220 pe, by BD (3 mentions) Cd34 fitc, by Thermo Fisher Scientific (2 mentions) Gr1 pe, by BD (2 mentions) Streptavidin pe secondary antibody, by BD (2 mentions) Facscalibur, by BD (2 mentions) Ter119 apc, by BD (2 mentions) Cd3 pe, by BD (2 mentions) Sca 1 pe, by BD (2 mentions) Lsrii flow cytometer, by BD (2 mentions) Cd45 apc, by BioLegend (2 mentions) Vimentin, by Merck Group (2 mentions) Cd31 apc, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

Anti-c-kit APC (7 citations) APC-conjugated anti-c-kit (4 citations) APC-conjugated c-kit (3 citations) C-kit APC-Cy7 (3 citations) C-Kit-APC (2B8), (3 citations) Alexa Fluor APC-conjugated antibody against human C-Kit (2 citations) Anti-c-Kit-APC (2B8)-APC (2 citations) C-Kit-APC-eFlour780 (2B8), (2 citations) APC-conjugated anti mouse CD117 (c-Kit) (2 citations) APC-conjugated anti-c-Kit (2B8, (2 citations)

24 protocols using c kit apc

Multiparametric Flow Cytometry Profiling of Hematopoietic Lineages

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Blood lymphoid. CD4-Fitc (BioLegend), CD8-PE-Cy7 (BD), B220–Pacific blue (BD), CD11b-PerCpCy5,5 (BD), CD3-APC (BioLegend); Blood myeloid and erythroid: CD3-Fitc (BD), CD19-Fitc (BD), B220-Fitc (BD), Gr1-PE (BD), F4/80-Pacific blue (BioLegend), CD11b-APC (BD);
BM lymphoid. IgD-Fitc (BD), CD25-PE (BioLegend), CD11b-PerCpCy5,5 (BD), IgM-PECy7 (Southern Biotech), B220-Pacific blue (BD), cKit-APC (BD); BM myeloid: Gr1-PE (BD), CD11b-PerCpCy5,5 (BD), Ter119-PECy7 (BD), F4/80–Pacific blue (BioLegend), CD19-APC (BioLegend); BM megakaryocyte: CD3-Fitc (BD), CD41-PE (BioLegend), CD11b-PerCpCy5,5 (BD), B220–Pacific blue (BD), CD19-APC (BioLegend); HSPC and CLP: Strep-APC/CY7 (Southern Biotech), cKit-APC (BD), SCA1-PacBlue (BioLegend), CD34-FITC (eBioscience), CD135-PE (BioLegend), CD150-PECy7 (BioLegend), CD127-PErCpCy5,5 (BioLegend); Hematopoietic progenitors: Strep-APC/CY7 (Southern Biotech), cKit-APC (BD), SCA1-PacBlue (BioLegend), CD34-FITC (eBioscience), CD16/32-PerCpCy5,5 (eBioscience), CD127-PE (eBioscience).

Alendar A., Lambooij J.P., Bhaskaran R., Lancini C., Song J.Y., van Vugt H., Snoek M, & Berns A. (2020). Gene expression regulation by the Chromodomain helicase DNA-binding protein 9 (CHD9) chromatin remodeler is dispensable for murine development. PLoS ONE, 15(5), e0233394.

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Isolation of Murine Hematopoietic Stem Cells

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Bone marrow cells were harvested from tibia and femurs of mice by gentle flushing with HBSS++ buffer [Hanks balanced salt solution (10–547F, Lonza) + HEPES (BP299–100, Fisher Scientific) + Fetal bovine serum (F2442, Sigma) + penicillin-streptomycin (15140–122, GIBCO)]. Samples were filtered through a 70 μM filter to obtain a single cell suspension and Lin⁻ enrichment was performed with lineage negative selection using the lineage cell depletion kit (130–090-858, Miltenyi). For isolation of LSK cells and LT-HSCs, Lin⁻ cells were incubated with biotin-labeled lineage antibody cocktail from BD containing a mixture of antibodies against CD3, CD11b, CD19, B220, Gr-1 and Ter119 (51–09082J, BD Pharmingen), followed by staining with streptavidin-PE secondary antibody (554061, BD). LSK cells were identified as Lin-Sca-1+c-kit+ using PE-Cy7-Sca (Clone D7, 558162, BD Biosciences) and APC-c-kit (Clone ACK2, 135108, BD Biosciences) antibodies; whereas LT-HSCs were recognized by being LSK and CD150+CD48- using PE-Cy7-Sca (Clone D7, 558162, BD Biosciences), APC-c-kit (Clone ACK2, 135108, BD Biosciences), Pacific Blue-CD150 (Clone TC15–12F12.2, 115924, Biolegend) and APC-Cy7-CD48 (Clone HM48–1, 47–0481-82, e-Bioscience). Cells were sorted using a BD FACSAria cell sorter.

Rodríguez A., Yang C., Furutani E., de Teresa B.G., Velazquez M., Filiatrault J., Sambel L.A., Phan T., Flores-Guzmán P., Sánchez S., Orozco A.M., Mayani H., Bolukbasi O.V., Färkkilä A., Epperly M., Greenberger J., Shimamura A., Frías S., Grompe M., Parmar K, & D’Andrea A.D. (2020). Inhibition of TGFβ1 and TGFβ3 promotes hematopoiesis in Fanconi anemia. Experimental hematology, 93, 70-84.e4.

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Multiparametric Analysis of Hematopoietic Cells

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PerCP-Cy 5.5 Mouse lineage cocktail, APC c-Kit, APC Gr-1, PE B220, APC anti-BrdU and Fc block (CD16/CD32) were all from BD Pharmingen. Pacific blue anti-mouse Ly-6A/E (Sca-1) was purchased from Biolegend. PE F4/80, APC CD11b and APC Annexin V were from eBioscience.

Maifrede S., Magimaidas A., Sha X., Mukherjee K., Liebermann D.A, & Hoffman B. (2017). Loss of Egr1, a human del5q gene, accelerates BCR-ABL driven chronic myelogenous leukemia. Oncotarget, 8(41), 69281-69294.

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Isolation of Long-Term Hematopoietic Stem Cells

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Mice were sacrificed and the bone marrow from tibia and femurs was harvested by gentle flushing with HBSS++ buffer [Hanks balanced salt solution (10-547F, Lonza) + HEPES (BP299-100, Fisher Scientific) + Fetal bovine serum (F2442, Sigma) + penicillin-streptomycin (15140–122, GIBCO)]. A 70 μM filter was used for filtering the samples and obtaining a single cell suspension. Lin^- enrichment was performed with lineage negative selection using the lineage cell depletion kit (130-090-858, Miltenyi).
For isolation of LT-HSCs, identified as Lin^-Sca-1⁺c-kit⁺ CD150⁺CD48^-, Lin^- cells were incubated with a mixture of biotin-labeled lineage antibody cocktail against CD3, CD11b, CD19, B220, Gr-1 and Ter119 (51-09082J, BD Pharmingen), and fluorochrome conjugate antibodies PE-Cy7-Sca (Clone D7, 558162, BD Biosciences), APC-c-kit (Clone ACK2, 135108, BD Biosciences), Pacific Blue-CD150 (Clone TC15-12F12.2, 115924, Biolegend) and APC-Cy7-CD48 (Clone HM48-1, 47-0481-82, e-Bioscience), followed by incubation with streptavidin-PE secondary antibody (554061, BD). LT-HSCs were sorted with a BD FACSAria cell sorter.

Rodríguez A., Epperly M., Filiatrault J., Velázquez M., Yang C., McQueen K., Sambel L.A., Nguyen H., Iyer D.R., Juárez U., Ayala-Zambrano C., Martignetti D.B., Frías S., Fisher R., Parmar K., Greenberger J.S, & D’Andrea A.D. (2022). TGFβ pathway is required for viable gestation of Fanconi anemia embryos. PLOS Genetics, 18(11), e1010459.

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Multiparameter Analysis of Spermatogonia

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Cells were digested into single cells using 0.05% trypsin and incubated in DMEM containing 10% fetal bovine serum (Thermo Fisher Scientific), stained with fluorochrome-conjugated antibodies, and washed with PBS before being analyzed, or sorted by a BD Fortessa or an Aria II flow cytometer (BD Biosciences). To determine the NAD+/NADH ratio, spermatogonia containing a SoNar probe were excited by 488 and 405 nm lasers, and fluorescence signals were detected through 530/30 FITC and 530/30 Alexa Fluor 430 channels, using BD Fortessa or Aria II flow cytometers. To determine ROS levels, cells were stained with 5 µM H₂DCFDA (D399, Thermo Fisher Scientific) in PBS containing 1% BSA at 37 °C for 40 min in the dark followed by washing with PBS twice before measurement, using flow cytometers. Antibodies were used with 2 µg/mL per 1–2 million cells as a final working concentration: APC-CD9 (17-0091-82, eBioscience, USA); APC-CD90.2 (105311), and FITC-c-Kit (105806) from Biolegend, USA; and PE-cy7-CD90.2 (561642), PE-c-Kit (553355), and APC-c-Kit (553356) from BD Bioscience, USA.

Chen W., Zhang Z., Chang C., Yang Z., Wang P., Fu H., Wei X., Chen E., Tan S., Huang W., Sun L., Ni T., Yang Y, & Wang Y. (2020). A bioenergetic shift is required for spermatogonial differentiation. Cell Discovery, 6, 56.

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Cell Cycle Analysis in Lymphoblast and HSC

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Cell cycle analysis in lymphoblast cell lines were thoroughly resuspended and fixed with ice-cold 70% ethanol and stored at −20 during 30 min. After fixation, cells were washed with PBS, stained with FxCycle PI/RNase staining solution (Invitrogen, F10797) and analyzed in a CytoFlex 5 Flow Cytometer (Beckman Coulter). Cell cycle analysis in mouse LT-HSC was performed using the cell surface markers PE-Cy7-Sca (Clone D7, 558162, BD Biosciences), APC-c-kit (Clone ACK2, 135108, BD Biosciences), FITC-CD150 (Clone A12 (7D4), 306306, Biolegend) and APC-Cy7-CD48 (Clone HM48-1, 470481-82, e-Bioscience), along with intracellular staining using PE-Cy7-Ki67 (350526, Biolegend) and DAPI (422801, Biolegend). Data analysis was performed using FlowJo software version 10.5.3.

Rodríguez A., Zhang K., Färkkilä A., Filiatrault J., Yang C., Velázquez M., Furutani E., Goldman D.C., de Teresa B.G., Garza-Mayen G., McQueen K., Sambel L.A., Molina B., Torres L., González M., Vadillo E., Pelayo R., Fleming W.H., Grompe M., Shimamura A., Hautaniemi S., Greenberger J., Frías S., Parmar K, & D’Andrea A.D. (2020). MYC Promotes Bone Marrow Stem Cell Dysfunction in Fanconi Anemia. Cell stem cell, 28(1), 33-47.e8.

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Isolation and Identification of Mouse LT-HSCs

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BM cells were harvested from tibia and femurs of mice by gentle flushing with HBSS++ buffer [Hanks balanced salt solution (10-547F, Lonza) + HEPES (BP299-100, Fisher Scientific) + Fetal bovine serum (F2442, Sigma) + penicillin-streptomycin (15140-122, GIBCO)]. Samples were filtered through a 70 μM filter and Lin- enrichment was performed using the lineage cell depletion kit (130-090-858, Miltenyi). LT-HSCs were recognized by being LSK (Lin-Sca-1+c-Kit+) and CD150+CD48- using PE-Cy7-Sca (Clone D7, 558162, BD Biosciences), APC-c-Kit (Clone ACK2, 135108, BD Biosciences), Pacific Blue-CD150 (Clone TC15-12F12.2, 115924, Biolegend) and APC-Cy7-CD48 (Clone HM48-1, 47-0481-82, e-Bioscience).

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Quantifying Hematopoietic Stem Cell Clones

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Clone size calculations were determined as previously described [12] (link). After 10 days of culture, single HSC-derived clones were estimated to be small (50–5,000 cells), medium (5,000–20,000 cells), or large (≥20,000). No clones had fewer than 50 cells. Ten-day clones were stained with biotinylated lineage marker antibodies (Haematopoietic Progenitor Enrichment Cocktail, STEMCELL), c-Kit-APC (BD) and Sca-1-Pacific Blue (Biolegend). To enumerate cells, a defined number of fluorescent beads (Trucount Control Beads, BD) were added to each well, and each sample was backcalculated to the proportion of the total that were run through the cytometer. Small clones were not able to be assessed individually by flow cytometry and were pooled; the percentage of c-Kit⁺Sca⁺Lin⁻ (KSL) cells was greater than 95%. Flow cytometry was performed on an LSRII Fortessa (BD), and all data were analyzed using Flowjo 10.0.7 (Treestar, USA).

Schulte R., Wilson N.K., Prick J.C., Cossetti C., Maj M.K., Gottgens B, & Kent D.G. (2015). Index sorting resolves heterogeneous murine hematopoietic stem cell populations. Experimental Hematology, 43(9), 803-811.

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Multiparametric Flow Cytometry Analysis of Hematopoietic Cells

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BM cells or splenocytes were harvested and subjected to red blood cell lysis. Fresh or frozen cells were stained with the following Abs: CD45.2-FITC and CD45.1-APC, Mac1-PE, Gr1-APC, c-Kit–APC, CD71-PE, Ter119-APC, B220-PE, and CD3-APC (BD) and analyzed on the BD FACSCalibur instrument. Staining for multiparameter flow cytometry was performed after a c-kit enrichment using 10 µl MACS beads (CD117) per mouse and then run on an AutoMACS (Miltenyi Biotec) according to the manufacturer’s instructions. The cells were then stained with the following cocktail: (Lineage; CD3, CD4, CD8, Gr1, B220, CD19, and TER119 all conjugated with PeCy5), Sca-Pac Blue, CD34-FITC or CD45.2-FITC, SLAM-APC, CD48-PE, c-KIT–Alexa Fluor 780, and FcgRIIb-PeCy7 (Fig. 2 d); HPCs (Lin^loc-Kit⁺Sca1⁻), GMPs (LK, FcγRIIb^hiCD34⁺), CMPs (LK, FcγRIIb^mid CD34⁺), MEPs (LK, FcγRIIb^loCD34⁻), B cells (B220⁺), and T cells (CD3⁺) from the spleen were also sorted. For analysis of LMPPs and CLPs, the following cocktail was used: Lineage marker mix–PeCy5, Sca-Pecy7 IL-7Ra–Pac Blue, Flk2-PE, CD34-FITC, and Kit-APC (Martin et al., 2003 (link); Adolfsson et al., 2005 (link); Karsunky et al., 2008 (link)).

Park S.M., Deering R.P., Lu Y., Tivnan P., Lianoglou S., Al-Shahrour F., Ebert B.L., Hacohen N., Leslie C., Daley G.Q., Lengner C.J, & Kharas M.G. (2014). Musashi-2 controls cell fate, lineage bias, and TGF-β signaling in HSCs. The Journal of Experimental Medicine, 211(1), 71-87.

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Hematopoietic Subpopulation Sorting

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Sorting of Lin-/kit+ hematopoietic subpopulations was conducted on a FACS Aria (BD Biosciences) and analyzed using FlowJo software (Flow Jo). Antibodies used to separate HSPC populations were as follows: streptavidin-PerCP-Cy5.5, c-kit-APC, FLT3-PE, and FcRγ-PE from BD Bioscience (San Jose, CA), and sca-1-PE-Cy7 and CD34-FITC from ebioscience (San Diego, CA).

Heiser D., Tan Y.S., Kaplan I., Godsey B., Morisot S., Cheng W.C., Small D, & Civin C.I. (2014). Correlated miR-mRNA Expression Signatures of Mouse Hematopoietic Stem and Progenitor Cell Subsets Predict “Stemness” and “Myeloid” Interaction Networks. PLoS ONE, 9(4), e94852.

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