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1) Product Images from "The eukaryotic signal sequence, YGRL, targets the chlamydial inclusion"
Article Title: The eukaryotic signal sequence, YGRL, targets the chlamydial inclusion
Journal: Frontiers in Cellular and Infection Microbiology
Figure Legend Snippet: Syntaxin 6 + YKGL or + YQRL protein-lipid interactions and localization to the chlamydial inclusion . In (A) , 3XFLAG-syntaxin 6 wild-type (WT), 3XFLAG-syntaxin 6 + YKGL (YKGL), and 3XFLAG-syntaxin 6 +YQRL (YQRL) were immunoprecipitated from HeLa cells and incubated with Sphingostrips or PIPstrips. Fold increase in syntaxin 6-lipid binding was determined by normalization of densitometry to input. Graph depicts the fold changes in densitometry levels of WT compared to YKGL or YQRL, respectively. The graph combines three independent Sphingostrip and PIPstrip assays. Averages and standard error of the mean are shown. In (B) , HeLa cells were transfected with WT, YKGL, or YQRL constructs, infected and fixed in ethanol as described in Figure 3 . The samples were processed for indirect immunofluorescence to detect the 3XFLAG (red), the inclusion membrane (green), or organisms (blue). Images are representative of two independent experiments. White arrows indicate colocalization with the inclusion; bars, 10 μm.
Techniques Used: Immunoprecipitation, Incubation, Binding Assay, Transfection, Construct, Infection, Immunofluorescence
Figure Legend Snippet: Syntaxin 6 and syntaxin 6ΔYGRL protein-lipid interactions . 3XFLAG empty vector (V), 3XFLAG-syntaxin 6 (Stx6) and 3XFLAG-syntaxin 6ΔYGRL (Stx6Δ) were immunopreciptated from HeLa cells and incubated with Sphingostrips (A) or PIPstrips (C) . Protein-lipid interactions with sulfatide (1), phosphatidylinositol 3-phosphate (PI3P) (2), phosphatidylinositol 4-phosphate (PI4P) (3), phosphatidylinositol 5-phosphate (PI5P) (4), and phosphatidylserine (PS) (5) were detected by blotting with anti-3XFLAG. The blank (Bl) is illuminated for orientation purposes. Protein input for sphingostrips and PIP strips was analyzed by blotting with anti-FLAG M2 antibody ( B,D , respectively). Fold increase in syntaxin 6-lipid binding was determined by normalization of densitometry to input (E) . Graph depicts the fold changes in densitometry levels of Stx6 compared to Stx6ΔYGRL. Positive reactions on Sphingostrips and PIPstrips were normalized to input and the graph combines data from three independent Sphingostrip and four independent PIPstrip assays. Averages and standard error of the mean are shown.
Techniques Used: Plasmid Preparation, Incubation, Binding Assay