Keratinocytes and hair follicle stem cells were isolated from the mouse dorsal skin as described previously (Xiong et al., 2022b ). Briefly, isolated dorsal skin samples were trypsinized, the epidermis was scrapped, minced, and filtered through a 70-µm cell strainer to prepare primary keratinocytes single-cell suspension. To isolate HFSC, the single-cell suspension was incubated with CD34-FITC antibody (eBioscience, 11-0341-82), Alexa 647-conjugated CD49f antibody (BD Biosciences, 562494), 7-AAD (BD Biosciences, 559925), and different Fluorescence minus one was used as a control. Cells then were sorted by BD FACS Aria and analyzed by Flow Jo.
Alexa 647 conjugated cd49f antibody
Manufactured by BD
The Alexa Fluor 647-conjugated CD49f antibody is a laboratory reagent used to detect and quantify the expression of the CD49f (α6 integrin) cell surface marker. CD49f is a component of the laminin receptor and is commonly used as a marker for stem cells and certain types of progenitor cells. This antibody is conjugated to the Alexa Fluor 647 fluorescent dye, which allows for detection and analysis of CD49f-positive cells using flow cytometry or other fluorescence-based techniques.
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2 protocols using alexa 647 conjugated cd49f antibody
1
Isolation of Mouse Keratinocytes and HFSC
2
Isolation and Characterization of Mouse and Human Hair Follicle Stem Cells
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Mouse keratinocytes and HFSCs were isolated from the mouse dorsal skin as described previously (Xiong et al., 2022b (link)). Briefly, isolated dorsal skin samples were trypsinized, the epidermis was scrapped, minced, and filtered through a 70 µm cell strainer to prepare primary keratinocytes single-cell suspension. To isolate HFSC, the single-cell suspension was incubated with CD34-FITC antibody (eBioscience, 11-0341-82), Alexa 647-conjugated CD49f antibody (BD Biosciences, 562494), 7-AAD (BD Biosciences, 559925), and different fluorescence minus one was used as a control. Cells were then sorted by BD FACS Aria and analyzed by Flow Jo. All the primary cells were used within 48 hr for experiments.
Human HFDPCs, mycoplasma tested, were purchased from Cell Applications, Inc (cat.# 602-05a). Human HFSCs, mycoplasma tested, were purchased from Celprogen (cat.# 36007-08). Human epidermal keratinocytes, neonatal, pooled, mycoplasma tested, were purchased from Lonza Reagents (cat.# 192906).
Human HFDPCs, mycoplasma tested, were purchased from Cell Applications, Inc (cat.# 602-05a). Human HFSCs, mycoplasma tested, were purchased from Celprogen (cat.# 36007-08). Human epidermal keratinocytes, neonatal, pooled, mycoplasma tested, were purchased from Lonza Reagents (cat.# 192906).
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