HMEC-1 WT cells were transfected with pCMV6-Entry (C-terminal Myc and DDK Tagged, OriGene Technologies, Rockville, MD) or OPN4 (Myc-DDK-Tagged)-pCMV6-Entry transcript variant 1 (TrueORF Gold Expression-Validated cDNA Clones from OriGene Technologies, Rockville, MD) using FuGene HD Transfection Reagent (Promega, Madison, WI). After transfection, cells were kept in complete darkness until room light (~200 LUX) or intense light (~10,000 LUX) exposures for 30 minutes. Melanopsin protein expression validation was done by isolating protein using RIPA buffer (Thermo Fisher Scientific, Waltham, MA) supplemented with protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA). Immunoblotting for anti-DDK (OriGene Technologies, Rockville, MD) was used to detect DDK-tagged melanopsin in transfected cells. Light-sensing cells subjected to glycolytic or mitochondrial stress tests on the Seahorse Bioanalyzer were exposed to light 30 min prior to Seahorse analyses.
Immunoblotting for anti ddk
Manufactured by OriGene
The Immunoblotting for anti-DDK is a laboratory equipment product. It is used for the detection and analysis of proteins tagged with the DDK (FLAG) epitope. The product provides the necessary tools and reagents to perform immunoblotting, a technique used to identify and quantify specific proteins in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (4 mentions)
Protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Pcmv6 entry c terminal myc and ddk tagged, by OriGene (2 mentions)
Myc ddk tagged pcmv6 entry, by OriGene (2 mentions)
Trueorf gold expression validated cdna clones, by OriGene (2 mentions)
Fugene hd transfection reagent, by Promega (2 mentions)
2 protocols using immunoblotting for anti ddk
1
Photoreceptive Cell Characterization
2
Photoreceptive Cell Characterization
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HMEC-1 WT cells were transfected with pCMV6-Entry (C-terminal Myc and DDK Tagged, OriGene Technologies, Rockville, MD) or OPN4 (Myc-DDK-Tagged)-pCMV6-Entry transcript variant 1 (TrueORF Gold Expression-Validated cDNA Clones from OriGene Technologies, Rockville, MD) using FuGene HD Transfection Reagent (Promega, Madison, WI). After transfection, cells were kept in complete darkness until room light (~200 LUX) or intense light (~10,000 LUX) exposures for 30 minutes. Melanopsin protein expression validation was done by isolating protein using RIPA buffer (Thermo Fisher Scientific, Waltham, MA) supplemented with protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA). Immunoblotting for anti-DDK (OriGene Technologies, Rockville, MD) was used to detect DDK-tagged melanopsin in transfected cells. Light-sensing cells subjected to glycolytic or mitochondrial stress tests on the Seahorse Bioanalyzer were exposed to light 30 min prior to Seahorse analyses.
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