48 hours after seeding at clonal density, NFSK were cultured with 50μM O-propargyl-puromycin (Jena Bioscience, NU-931-05) in keratinocyte media for 1 hour. Plates were then fixed with 4% paraformaldehyde for 10 minutes on ice. Colonies were stained following the same protocol described above for immnostaining with EdU using a 1:500 dilution of a Keratin 1 antibody (AF-87, Covance) and an Alexa Fluor Click-IT kit (Invitrogen, C10337) to detect O-propargyl-puromycin. Colonies containing 6 to 10 cells were imaged using a Zeiss Axio-Observer D1 microscope with Zeiss Axiovision software and mean colony fluorescence quantified using NIH ImageJ software.
Alexa fluor click it kit
Manufactured by Thermo Fisher Scientific
The Alexa Fluor Click-IT kit is a laboratory product designed for the detection and analysis of biomolecules. The kit utilizes a Click chemistry-based approach to enable the labeling and visualization of target molecules. It provides a versatile tool for researchers in various fields of study.
Automatically generated - may contain errors
2 protocols using alexa fluor click it kit
1
Measuring Cell Proliferation in Keratinocytes
2
Measuring Cell Proliferation in Keratinocytes
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48 hours after seeding at clonal density, NFSK were cultured with 50μM O-propargyl-puromycin (Jena Bioscience, NU-931-05) in keratinocyte media for 1 hour. Plates were then fixed with 4% paraformaldehyde for 10 minutes on ice. Colonies were stained following the same protocol described above for immnostaining with EdU using a 1:500 dilution of a Keratin 1 antibody (AF-87, Covance) and an Alexa Fluor Click-IT kit (Invitrogen, C10337) to detect O-propargyl-puromycin. Colonies containing 6 to 10 cells were imaged using a Zeiss Axio-Observer D1 microscope with Zeiss Axiovision software and mean colony fluorescence quantified using NIH ImageJ software.
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