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Cd140a

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CD140a is a cell surface receptor that functions as a transmembrane protein. It is involved in the regulation of various cellular processes.

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Lab products found in correlation

Foxp3 fjk 16s, by Thermo Fisher Scientific (1 mentions) Biotinylated ulex europaeus agglutinin 1 uea 1, by Vector Laboratories (1 mentions) Cd11b clone m1 70, by Thermo Fisher Scientific (1 mentions) Foxp3 fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions) Cd105, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Cd11b, by Thermo Fisher Scientific (1 mentions) Ly 6a ly 6e sca1, by BD (1 mentions) Fab3518n, by Thermo Fisher Scientific (1 mentions) Dispase 2, by Roche (1 mentions) Collagenase b, by Roche (1 mentions)

4 protocols using cd140a

1

Isolation and Characterization of Mouse Skin Cell Populations

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Mouse dorsal skin was collected, and the fat layer was removed by gentle scrapping from the dermal side. The skin was incubated in 0.25% collagenase in HBSS at 37 °C for 35–45 minutes on an orbital shaker. Single cell suspension was collected by gentle scraping of the dermal side and filtering through 70 μm and 40 μm filters. The epidermal layer was incubated in trypsin-EDTA at 37 °C for 35–45 minutes on an orbital shaker. Single cell suspension was collected by gentle scraping of the epidermal side and filtering through 70 μm and 40 μm filters. The single cell suspension was centrifuged for 5 minutes at 4°C, resuspended in 0.25% FBS in PBS, and stained with fluorescent dye-conjugated antibodies for 30 minutes. For late anagen skin samples, the bottom parts of the hair follicles containing mature melanocytes were removed by gentle scrapping under dissection microscope. The MeSCs located close to the bulge remained and were verified by immunostaining. Antibodies used: CD140a (Invitrogen 13–1401-82, 1:200), CD45 (Invitrogen 13–0451-82, 1:400), Sca1 (Invitrogen 13–5981-82, 1:1000), CD34 (Invitrogen 13–0341-82, 1:100), CD117 (Biolegend 135136, Dilution 1:400). See a protocol published at protocol exchange website for a step-to-step instruction57 (link).
Zhang B., Ma S., Rachmin I., He M., Baral P., Choi S., Gonçalves W.A., Shwartz Y., Fast E.M., Su Y., Zon L.I., Regev A., Buenrostro J.D., Cunha T.M., Chiu I.M., Fisher D.E, & Hsu Y.C. (2020). Hyperactivation of Sympathetic Nerves Drives Melanocyte Stem Cell Depletion. Nature, 577(7792), 676-681.
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2

Isolation and Characterization of Mouse Skin Cell Populations

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Mouse dorsal skin was collected, and the fat layer was removed by gentle scrapping from the dermal side. The skin was incubated in 0.25% collagenase in HBSS at 37 °C for 35–45 minutes on an orbital shaker. Single cell suspension was collected by gentle scraping of the dermal side and filtering through 70 μm and 40 μm filters. The epidermal layer was incubated in trypsin-EDTA at 37 °C for 35–45 minutes on an orbital shaker. Single cell suspension was collected by gentle scraping of the epidermal side and filtering through 70 μm and 40 μm filters. The single cell suspension was centrifuged for 5 minutes at 4°C, resuspended in 0.25% FBS in PBS, and stained with fluorescent dye-conjugated antibodies for 30 minutes. For late anagen skin samples, the bottom parts of the hair follicles containing mature melanocytes were removed by gentle scrapping under dissection microscope. The MeSCs located close to the bulge remained and were verified by immunostaining. Antibodies used: CD140a (Invitrogen 13–1401-82, 1:200), CD45 (Invitrogen 13–0451-82, 1:400), Sca1 (Invitrogen 13–5981-82, 1:1000), CD34 (Invitrogen 13–0341-82, 1:100), CD117 (Biolegend 135136, Dilution 1:400). See a protocol published at protocol exchange website for a step-to-step instruction57 (link).
Zhang B., Ma S., Rachmin I., He M., Baral P., Choi S., Gonçalves W.A., Shwartz Y., Fast E.M., Su Y., Zon L.I., Regev A., Buenrostro J.D., Cunha T.M., Chiu I.M., Fisher D.E, & Hsu Y.C. (2020). Hyperactivation of Sympathetic Nerves Drives Melanocyte Stem Cell Depletion. Nature, 577(7792), 676-681.
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3

Multiparameter Flow Cytometry Analysis

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The antibodies and reagents used for flow cytometry analysis are listed: anti-mouse antibodies, CD11b (clone M1/70), CD29 (HMb1-1), CD31 (390), CD34 (RAM34), CD44 (IM7), CD45 (30-F11), CD105 (MJ7/18), CD106 (429), CD140a (APA5), Sca-1 (Ly-6A/E, D7), CD3(17A2), CD4 (GK1.5), CD8a (53-6.7), CD25 (PC61.5), FoxP3 (FJK-16s), T-cell receptor Vβ 5.1/5.2 (MR9-4), CD326 (EpCAM, G8.8), Ly51(BP-1, 6C3), Aire (5H12), CCR9 (CD199, CW-1.2), and allophycocyanin-labeled streptavidin (17-4317-82), which were obtained from eBioscience. Biotinylated Ulex europaeus agglutinin-1 (UEA-1, #B1065) was purchased from Vector Laboratories. The Foxp3 permeabilization/fixation kit (eBioscience) was used for cell fixation/permeabilization during intracellular staining. Aqua fluorescent reactive dye for viability analysis (L34957) was obtained from Invitrogen. All staining was performed per the manufacturers’ instructions. Labeled cells were analyzed on a BD FACS Canto II (BD Biosciences, Franklin Lakes, NJ) and data analyzed using the FlowJo software. The gating strategies are shown in the supplemental Figure 6.
Zhang X., He J., Zhao K., Liu S., Xuan L., Chen S., Xue R., Lin R., Xu J., Zhang Y., Xiang A.P., Jin H, & Liu Q. (2023). Mesenchymal stromal cells ameliorate chronic GVHD by boosting thymic regeneration in a CCR9-dependent manner in mice. Blood Advances, 7(18), 5359-5373.
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4

Isolating Muscle Stem Cells from Mice

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For the isolation of muscle stem cells, uninjured hindlimb muscles representing various fiber types (gastrocnemius/soleus/plantaris complex, tibialis anterior/EDL complex and quadriceps) were collected and from PpargΔ/Δ mice and their control littermates and freshly digested with dispase II (2.5 U/mL) (Roche, Basel, Switzerland), collagenase B (0.2%) (Roche), and MgCl2 (5 mM). Cells were then incubated at 4 °C for 30 min with fluorescently-coupled antibodies against CD45 (Invitrogen, Carlsbad, CA, USA) MCD4501 or MCD4528; dilution for both 1/25), CD31 (Invitrogen, RM5201 or RM5228; dilution for both 1/25), CD11b (Invitrogen, RM2801 or RM2828; dilution for both 1/25), CD34 (BD Biosciences, 560230 or 560238; dilution for both 1/60), Ly-6A–Ly-6E (Sca1) (BD Biosciences, Franklin Lakes, NJ, USA) 561021; dilution 1/150), α7-integrin (R&D, FAB3518N; dilution 1/30) and CD140a (eBioscience, San Diego, CA, USA, 12-1401-81 or 17-1401-81; dilution for both 1/30). Cell isolation was performed on a Beckman–Coulter Astrios Cell sorter; Beckman-Coulter, Brea, CA, USA. MuSCs isolated by flow-cytometry were CD45−CD31−CD11b−Sca1−CD34+Integrinα7+; fibro/adipogenic progenitors (FAPs) were CD45−CD31−CD11b−Sca1+CD34+PDGFRα+.
Dammone G., Karaz S., Lukjanenko L., Winkler C., Sizzano F., Jacot G., Migliavacca E., Palini A., Desvergne B., Gilardi F, & Feige J.N. (2018). PPARγ Controls Ectopic Adipogenesis and Cross-Talks with Myogenesis During Skeletal Muscle Regeneration. International Journal of Molecular Sciences, 19(7), 2044.
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