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calmodulin antagonist  (MedChemExpress)
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MedChemExpress calmodulin antagonist
( A ) Design of pDXA-GCaMP3 plasmids. A Dictyostelium -specific calcium reporter plasmid, pDXA-GCaMP3, was developed by subcloning the GCaMP region (1,348 bp) from the mammalian expression plasmid pGCaMP3 (15) into the BamH1 site of the Dictyostelium expression vector pDXA-3C (6,113 bp). ( B ) Schematic of fluorescence expression upon calcium binding. GCaMP consists of a calcium-binding protein, <t>calmodulin</t> (CaM), a myosin light chain kinase (M13), and green fluorescent protein. Calcium-bound CaM interacts with M13, resulting in fluorescence. ( C ) Confirmation of GCaMP3 expression in Dictyostelium cells. The calcium reporter plasmid was introduced into wild-type (KAx3) and mutant cells. GCaMP3 expression was confirmed by immunoblotting with anti-GFP antibodies. Wild-type cells expressing empty GFP were used as a control in the first lane. ( D ) Fluorescence in vegetative Dictyostelium cells. Arrow heads indicate the protruding, ruffled regions of the cell. Scale bar = 5 μm.
Calmodulin Antagonist, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) Design of pDXA-GCaMP3 plasmids. A Dictyostelium -specific calcium reporter plasmid, pDXA-GCaMP3, was developed by subcloning the GCaMP region (1,348 bp) from the mammalian expression plasmid pGCaMP3 (15) into the BamH1 site of the Dictyostelium expression vector pDXA-3C (6,113 bp). ( B ) Schematic of fluorescence expression upon calcium binding. GCaMP consists of a calcium-binding protein, calmodulin (CaM), a myosin light chain kinase (M13), and green fluorescent protein. Calcium-bound CaM interacts with M13, resulting in fluorescence. ( C ) Confirmation of GCaMP3 expression in Dictyostelium cells. The calcium reporter plasmid was introduced into wild-type (KAx3) and mutant cells. GCaMP3 expression was confirmed by immunoblotting with anti-GFP antibodies. Wild-type cells expressing empty GFP were used as a control in the first lane. ( D ) Fluorescence in vegetative Dictyostelium cells. Arrow heads indicate the protruding, ruffled regions of the cell. Scale bar = 5 μm.

Journal: Journal of Microbiology and Biotechnology

Article Title: Intracellular Calcium Responses to External Calcium Stimuli in Dictyostelium

doi: 10.4014/jmb.2412.12066

Figure Lengend Snippet: ( A ) Design of pDXA-GCaMP3 plasmids. A Dictyostelium -specific calcium reporter plasmid, pDXA-GCaMP3, was developed by subcloning the GCaMP region (1,348 bp) from the mammalian expression plasmid pGCaMP3 (15) into the BamH1 site of the Dictyostelium expression vector pDXA-3C (6,113 bp). ( B ) Schematic of fluorescence expression upon calcium binding. GCaMP consists of a calcium-binding protein, calmodulin (CaM), a myosin light chain kinase (M13), and green fluorescent protein. Calcium-bound CaM interacts with M13, resulting in fluorescence. ( C ) Confirmation of GCaMP3 expression in Dictyostelium cells. The calcium reporter plasmid was introduced into wild-type (KAx3) and mutant cells. GCaMP3 expression was confirmed by immunoblotting with anti-GFP antibodies. Wild-type cells expressing empty GFP were used as a control in the first lane. ( D ) Fluorescence in vegetative Dictyostelium cells. Arrow heads indicate the protruding, ruffled regions of the cell. Scale bar = 5 μm.

Article Snippet: A calmodulin antagonist, W7, and 2,5-Di-tert-butylhydroquinone (BHQ) were purchased from MedChemExpress (USA).

Techniques: Plasmid Preparation, Subcloning, Expressing, Fluorescence, Binding Assay, Mutagenesis, Western Blot, Control