Structured Review

R&D Systems hek293 derived native human shh n protein
Functional validation of DISP1* for structural investigation. (A) A diagram of topology of human DISP1. Residues 1–171 and 1,249–1,524 were removed in DISP1*. (B) Experimental scheme to measure SHH release capacity. <t>HEK293</t> Flp-In T-REX cells were stably transfected with an inducible 3× Flag-tagged Scube2. Cells were induced and the supernatant containing Scube2 conditioned medium was collected, filtered, and mixed with fresh medium at a 1:1 ratio and finally transferred to DISP1 −/− or DISP1* and DISP FL rescued MEF cells stably expressing FL-SHH. After 48 h of incubation, the SHH-N conditioned medium was harvested and mixed with fresh medium at 1:1 ratio to incubate with SHH-Light II cells. (C) SHH-N release in DISP1 −/− MEF cells that were transduced with both SHH and empty vector (DISP1 KO ), full-length DISP1 (DISP1 FL ) or DISP1* was checked via dual luciferase assay where the SHH-Light II cells stably express firefly luciferase with an 8×-Gli promoter and Renilla luciferase with a constitutive promoter. Data are mean ± SD (n = 8 biologically independent experiments). (D) Pull-down assay of native SHH-N with DISP1*-WT or DISP1*–NNN mutant detected by Coomassie staining.
Hek293 Derived Native Human Shh N Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hek293 derived native human shh n protein - by Bioz Stars, 2021-01
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1) Product Images from "Structure of human Dispatched-1 provides insights into Hedgehog ligand biogenesis"

Article Title: Structure of human Dispatched-1 provides insights into Hedgehog ligand biogenesis

Journal: Life Science Alliance

doi: 10.26508/lsa.202000776

Functional validation of DISP1* for structural investigation. (A) A diagram of topology of human DISP1. Residues 1–171 and 1,249–1,524 were removed in DISP1*. (B) Experimental scheme to measure SHH release capacity. HEK293 Flp-In T-REX cells were stably transfected with an inducible 3× Flag-tagged Scube2. Cells were induced and the supernatant containing Scube2 conditioned medium was collected, filtered, and mixed with fresh medium at a 1:1 ratio and finally transferred to DISP1 −/− or DISP1* and DISP FL rescued MEF cells stably expressing FL-SHH. After 48 h of incubation, the SHH-N conditioned medium was harvested and mixed with fresh medium at 1:1 ratio to incubate with SHH-Light II cells. (C) SHH-N release in DISP1 −/− MEF cells that were transduced with both SHH and empty vector (DISP1 KO ), full-length DISP1 (DISP1 FL ) or DISP1* was checked via dual luciferase assay where the SHH-Light II cells stably express firefly luciferase with an 8×-Gli promoter and Renilla luciferase with a constitutive promoter. Data are mean ± SD (n = 8 biologically independent experiments). (D) Pull-down assay of native SHH-N with DISP1*-WT or DISP1*–NNN mutant detected by Coomassie staining.
Figure Legend Snippet: Functional validation of DISP1* for structural investigation. (A) A diagram of topology of human DISP1. Residues 1–171 and 1,249–1,524 were removed in DISP1*. (B) Experimental scheme to measure SHH release capacity. HEK293 Flp-In T-REX cells were stably transfected with an inducible 3× Flag-tagged Scube2. Cells were induced and the supernatant containing Scube2 conditioned medium was collected, filtered, and mixed with fresh medium at a 1:1 ratio and finally transferred to DISP1 −/− or DISP1* and DISP FL rescued MEF cells stably expressing FL-SHH. After 48 h of incubation, the SHH-N conditioned medium was harvested and mixed with fresh medium at 1:1 ratio to incubate with SHH-Light II cells. (C) SHH-N release in DISP1 −/− MEF cells that were transduced with both SHH and empty vector (DISP1 KO ), full-length DISP1 (DISP1 FL ) or DISP1* was checked via dual luciferase assay where the SHH-Light II cells stably express firefly luciferase with an 8×-Gli promoter and Renilla luciferase with a constitutive promoter. Data are mean ± SD (n = 8 biologically independent experiments). (D) Pull-down assay of native SHH-N with DISP1*-WT or DISP1*–NNN mutant detected by Coomassie staining.

Techniques Used: Functional Assay, Stable Transfection, Transfection, Expressing, Incubation, Transduction, Plasmid Preparation, Luciferase, Pull Down Assay, Mutagenesis, Staining

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    R&D Systems human disp1 antibody
    Human Disp1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human disp1 antibody/product/R&D Systems
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human disp1 antibody - by Bioz Stars, 2021-01
    90/100 stars
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