Review



stim1 encoding sequence  (Promega)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 90
      Buy from Supplier

    Structured Review

    Promega stim1 encoding sequence
    Functional analysis of the N-FLAG-tagged Orai1 channel. A, thapsigargin (TG)-induced [Ca2+]i rises in Orai1-expressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 (gray triangles)-, N-FLAG-tagged Orai1 (black circles)-, and control vector (white circles)-transfected HEK293 cells (n = 27–28). The perfusion solution was first changed to 0.5 mm EGTA containing Ca2+-free HBS (EGTA), and 2 μm TG was applied to the cells in the absence of extracellular Ca2+. Eleven minutes after the application of TG, 2 mm Ca2+ was added to the extracellular solution. Right, maximum [Ca2+]i rises (Δ[Ca2+]i) induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. B, TG-induced [Ca2+]i rises in Orai1 and <t>STIM1-coexpressing</t> HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 plus STIM1 (gray triangles)- and N-FLAG-tagged Orai1 plus STIM1 (black circles)-cotransfected HEK293 cells and control vector (white circles)-transfected HEK293 cells (n = 25–41). Right, Δ[Ca2+]i induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. C, activation of CRAC current induced by 40 μm IP3 in Orai1- and STIM1-coexpressing cells. Left, representative current-voltage relationships in Orai1 plus STIM1 (a)- and N-FLAG-tagged Orai1 plus STIM1 (b)-cotransfected HEK293 cells and control vector (c)-transfected HEK293 cells. Data represent leak-subtracted current densities (pA/picofarad) evoked by 50-ms voltage ramps from -100 to +100 mV. Right, average CRAC current densities at -80 mV in Orai1 plus STIM1 (n = 11), N-FLAG-tagged Orai1 plus STIM1 (n = 9) and control (n = 9) cells. **, p < 0.01; and ***, p < 0.001.
    Stim1 Encoding Sequence, supplied by Promega, 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/product/stim1+encoding+sequence/plasmid+vectors+of+the+stim1+promoter+and+the+promoter+with+a1++a2++or+a3+deletion/pmc02679469-38-15-11
    Average 90 stars, based on 1 article reviews
    stim1 encoding sequence - by Bioz Stars, 2026-11
    90/100 stars

    Images

    1) Product Images from "Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain * "

    Article Title: Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain *

    Journal:

    doi: 10.1074/jbc.M900812200

    Functional analysis of the N-FLAG-tagged Orai1 channel. A, thapsigargin (TG)-induced [Ca2+]i rises in Orai1-expressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 (gray triangles)-, N-FLAG-tagged Orai1 (black circles)-, and control vector (white circles)-transfected HEK293 cells (n = 27–28). The perfusion solution was first changed to 0.5 mm EGTA containing Ca2+-free HBS (EGTA), and 2 μm TG was applied to the cells in the absence of extracellular Ca2+. Eleven minutes after the application of TG, 2 mm Ca2+ was added to the extracellular solution. Right, maximum [Ca2+]i rises (Δ[Ca2+]i) induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. B, TG-induced [Ca2+]i rises in Orai1 and STIM1-coexpressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 plus STIM1 (gray triangles)- and N-FLAG-tagged Orai1 plus STIM1 (black circles)-cotransfected HEK293 cells and control vector (white circles)-transfected HEK293 cells (n = 25–41). Right, Δ[Ca2+]i induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. C, activation of CRAC current induced by 40 μm IP3 in Orai1- and STIM1-coexpressing cells. Left, representative current-voltage relationships in Orai1 plus STIM1 (a)- and N-FLAG-tagged Orai1 plus STIM1 (b)-cotransfected HEK293 cells and control vector (c)-transfected HEK293 cells. Data represent leak-subtracted current densities (pA/picofarad) evoked by 50-ms voltage ramps from -100 to +100 mV. Right, average CRAC current densities at -80 mV in Orai1 plus STIM1 (n = 11), N-FLAG-tagged Orai1 plus STIM1 (n = 9) and control (n = 9) cells. **, p < 0.01; and ***, p < 0.001.
    Figure Legend Snippet: Functional analysis of the N-FLAG-tagged Orai1 channel. A, thapsigargin (TG)-induced [Ca2+]i rises in Orai1-expressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 (gray triangles)-, N-FLAG-tagged Orai1 (black circles)-, and control vector (white circles)-transfected HEK293 cells (n = 27–28). The perfusion solution was first changed to 0.5 mm EGTA containing Ca2+-free HBS (EGTA), and 2 μm TG was applied to the cells in the absence of extracellular Ca2+. Eleven minutes after the application of TG, 2 mm Ca2+ was added to the extracellular solution. Right, maximum [Ca2+]i rises (Δ[Ca2+]i) induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. B, TG-induced [Ca2+]i rises in Orai1 and STIM1-coexpressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 plus STIM1 (gray triangles)- and N-FLAG-tagged Orai1 plus STIM1 (black circles)-cotransfected HEK293 cells and control vector (white circles)-transfected HEK293 cells (n = 25–41). Right, Δ[Ca2+]i induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. C, activation of CRAC current induced by 40 μm IP3 in Orai1- and STIM1-coexpressing cells. Left, representative current-voltage relationships in Orai1 plus STIM1 (a)- and N-FLAG-tagged Orai1 plus STIM1 (b)-cotransfected HEK293 cells and control vector (c)-transfected HEK293 cells. Data represent leak-subtracted current densities (pA/picofarad) evoked by 50-ms voltage ramps from -100 to +100 mV. Right, average CRAC current densities at -80 mV in Orai1 plus STIM1 (n = 11), N-FLAG-tagged Orai1 plus STIM1 (n = 9) and control (n = 9) cells. **, p < 0.01; and ***, p < 0.001.

    Techniques Used: Functional Assay, Expressing, Control, Plasmid Preparation, Transfection, Activation Assay

    Predicted topology of Orai1 and a docking model with STIM1. A, schematic diagram of N-FLAG Orai1. Orai1 is a highly glycosylated protein with glycan moiety of ∼8 kDa per subunit. The Orai1 polypeptide has a cytosolic N terminus, four TM segments (yellow columns: S1–S4), and a cytosolic C terminus with a putative coiled-coil sequence (green box) involved in binding with STIM1. The FLAG tag (red box) was introduced in the N terminus. B, illustration of putative docking between Orai1 and STIM1. The three-dimensional reconstruction in this study suggests that the cytoplasmic domain of Orai1 is long enough for direct association with STIM1. STIM1 contains a coiled-coil motif (green box) in the cytoplasm, a single TM segment (yellow column), a sterile α motif domain (purple box), and an EF-hand motif (blue crescent) in the ER lumen.
    Figure Legend Snippet: Predicted topology of Orai1 and a docking model with STIM1. A, schematic diagram of N-FLAG Orai1. Orai1 is a highly glycosylated protein with glycan moiety of ∼8 kDa per subunit. The Orai1 polypeptide has a cytosolic N terminus, four TM segments (yellow columns: S1–S4), and a cytosolic C terminus with a putative coiled-coil sequence (green box) involved in binding with STIM1. The FLAG tag (red box) was introduced in the N terminus. B, illustration of putative docking between Orai1 and STIM1. The three-dimensional reconstruction in this study suggests that the cytoplasmic domain of Orai1 is long enough for direct association with STIM1. STIM1 contains a coiled-coil motif (green box) in the cytoplasm, a single TM segment (yellow column), a sterile α motif domain (purple box), and an EF-hand motif (blue crescent) in the ER lumen.

    Techniques Used: Glycoproteomics, Sequencing, Binding Assay, FLAG-tag, Sterility

    Related Articles

    Functional Assay:

    Article Title: Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain
    Article Snippet: Expression Constructs and Transfection of Cells —Full-length mouse Orai1 encoding sequence (GenBankTM: {"type":"entrez-nucleotide","attrs":{"text":"NM_175423","term_id":"93277106","term_text":"NM_175423"}} NM_175423 ) was cloned from mouse brain Marathon-Ready cDNA (Clontech, Mountain View, CA) using PCR and was subcloned into pCMV-Tag2B or pCMV-Tag4B vector (Stratagene, La Jolla, CA) to introduce a FLAG tag sequence at the N or C terminus (N-FLAG or C-FLAG Orai1). .. For functional analysis, Orai1 encoding sequence was also subcloned into pCI-neo (Promega, Madison, WI), and mouse STIM1 encoding sequence was subcloned from pApuro-FLAG-tagged mouse STIM1 ( 31 ) into pCI-neo. ..

    Sequencing:

    Article Title: Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain
    Article Snippet: Expression Constructs and Transfection of Cells —Full-length mouse Orai1 encoding sequence (GenBankTM: {"type":"entrez-nucleotide","attrs":{"text":"NM_175423","term_id":"93277106","term_text":"NM_175423"}} NM_175423 ) was cloned from mouse brain Marathon-Ready cDNA (Clontech, Mountain View, CA) using PCR and was subcloned into pCMV-Tag2B or pCMV-Tag4B vector (Stratagene, La Jolla, CA) to introduce a FLAG tag sequence at the N or C terminus (N-FLAG or C-FLAG Orai1). .. For functional analysis, Orai1 encoding sequence was also subcloned into pCI-neo (Promega, Madison, WI), and mouse STIM1 encoding sequence was subcloned from pApuro-FLAG-tagged mouse STIM1 ( 31 ) into pCI-neo. ..

    other:

    Article Title: miR-195 competes with HuR to modulate stim1 mRNA stability and regulate cell migration
    Article Snippet: The full-length Stim1 5′-UTR, CR or 3′-UTR and different 3′-UTR fragments with or without predicted miR-195 binding sites were amplified and subcloned into the pmirGLO Dual-Luciferase miRNA Target Expression Vector (Promega, Madison, WI) to generate the pmirGLO-Luc-Stim1-5′UTR, pmirGLO-Luc-Stim1-CR and pmirGLO-Stim1-3′UTR.

    Plasmid Preparation:

    Article Title: STIM1 Mediates Hypoxia-Driven Hepatocarcinogenesis via Interaction with HIF-1.
    Article Snippet: .. Plasmid vectors of the STIM1 promoter and the promoter with A1, A2, or A3 deletion were synthesized by Promega (Madison) and transfected into HepG2 cells as described above. ..

    Synthesized:

    Article Title: STIM1 Mediates Hypoxia-Driven Hepatocarcinogenesis via Interaction with HIF-1.
    Article Snippet: .. Plasmid vectors of the STIM1 promoter and the promoter with A1, A2, or A3 deletion were synthesized by Promega (Madison) and transfected into HepG2 cells as described above. ..

    Transfection:

    Article Title: STIM1 Mediates Hypoxia-Driven Hepatocarcinogenesis via Interaction with HIF-1.
    Article Snippet: .. Plasmid vectors of the STIM1 promoter and the promoter with A1, A2, or A3 deletion were synthesized by Promega (Madison) and transfected into HepG2 cells as described above. ..



    Similar Products

    stim1 encoding sequence  (Promega)
    90
    Promega stim1 encoding sequence
    Functional analysis of the N-FLAG-tagged Orai1 channel. A, thapsigargin (TG)-induced [Ca2+]i rises in Orai1-expressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 (gray triangles)-, N-FLAG-tagged Orai1 (black circles)-, and control vector (white circles)-transfected HEK293 cells (n = 27–28). The perfusion solution was first changed to 0.5 mm EGTA containing Ca2+-free HBS (EGTA), and 2 μm TG was applied to the cells in the absence of extracellular Ca2+. Eleven minutes after the application of TG, 2 mm Ca2+ was added to the extracellular solution. Right, maximum [Ca2+]i rises (Δ[Ca2+]i) induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. B, TG-induced [Ca2+]i rises in Orai1 and <t>STIM1-coexpressing</t> HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 plus STIM1 (gray triangles)- and N-FLAG-tagged Orai1 plus STIM1 (black circles)-cotransfected HEK293 cells and control vector (white circles)-transfected HEK293 cells (n = 25–41). Right, Δ[Ca2+]i induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. C, activation of CRAC current induced by 40 μm IP3 in Orai1- and STIM1-coexpressing cells. Left, representative current-voltage relationships in Orai1 plus STIM1 (a)- and N-FLAG-tagged Orai1 plus STIM1 (b)-cotransfected HEK293 cells and control vector (c)-transfected HEK293 cells. Data represent leak-subtracted current densities (pA/picofarad) evoked by 50-ms voltage ramps from -100 to +100 mV. Right, average CRAC current densities at -80 mV in Orai1 plus STIM1 (n = 11), N-FLAG-tagged Orai1 plus STIM1 (n = 9) and control (n = 9) cells. **, p < 0.01; and ***, p < 0.001.
    Stim1 Encoding Sequence, supplied by Promega, 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/product/stim1+encoding+sequence/plasmid+vectors+of+the+stim1+promoter+and+the+promoter+with+a1++a2++or+a3+deletion/pmc02679469-38-15-11
    Average 90 stars, based on 1 article reviews
    stim1 encoding sequence - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Functional analysis of the N-FLAG-tagged Orai1 channel. A, thapsigargin (TG)-induced [Ca2+]i rises in Orai1-expressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 (gray triangles)-, N-FLAG-tagged Orai1 (black circles)-, and control vector (white circles)-transfected HEK293 cells (n = 27–28). The perfusion solution was first changed to 0.5 mm EGTA containing Ca2+-free HBS (EGTA), and 2 μm TG was applied to the cells in the absence of extracellular Ca2+. Eleven minutes after the application of TG, 2 mm Ca2+ was added to the extracellular solution. Right, maximum [Ca2+]i rises (Δ[Ca2+]i) induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. B, TG-induced [Ca2+]i rises in Orai1 and STIM1-coexpressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 plus STIM1 (gray triangles)- and N-FLAG-tagged Orai1 plus STIM1 (black circles)-cotransfected HEK293 cells and control vector (white circles)-transfected HEK293 cells (n = 25–41). Right, Δ[Ca2+]i induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. C, activation of CRAC current induced by 40 μm IP3 in Orai1- and STIM1-coexpressing cells. Left, representative current-voltage relationships in Orai1 plus STIM1 (a)- and N-FLAG-tagged Orai1 plus STIM1 (b)-cotransfected HEK293 cells and control vector (c)-transfected HEK293 cells. Data represent leak-subtracted current densities (pA/picofarad) evoked by 50-ms voltage ramps from -100 to +100 mV. Right, average CRAC current densities at -80 mV in Orai1 plus STIM1 (n = 11), N-FLAG-tagged Orai1 plus STIM1 (n = 9) and control (n = 9) cells. **, p < 0.01; and ***, p < 0.001.

    Journal:

    Article Title: Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain *

    doi: 10.1074/jbc.M900812200

    Figure Lengend Snippet: Functional analysis of the N-FLAG-tagged Orai1 channel. A, thapsigargin (TG)-induced [Ca2+]i rises in Orai1-expressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 (gray triangles)-, N-FLAG-tagged Orai1 (black circles)-, and control vector (white circles)-transfected HEK293 cells (n = 27–28). The perfusion solution was first changed to 0.5 mm EGTA containing Ca2+-free HBS (EGTA), and 2 μm TG was applied to the cells in the absence of extracellular Ca2+. Eleven minutes after the application of TG, 2 mm Ca2+ was added to the extracellular solution. Right, maximum [Ca2+]i rises (Δ[Ca2+]i) induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. B, TG-induced [Ca2+]i rises in Orai1 and STIM1-coexpressing HEK293 cells. Left, average time course of Ca2+ responses induced by 2 μm TG in Orai1 plus STIM1 (gray triangles)- and N-FLAG-tagged Orai1 plus STIM1 (black circles)-cotransfected HEK293 cells and control vector (white circles)-transfected HEK293 cells (n = 25–41). Right, Δ[Ca2+]i induced by TG in EGTA and those after re-addition of 2 mm external Ca2+. C, activation of CRAC current induced by 40 μm IP3 in Orai1- and STIM1-coexpressing cells. Left, representative current-voltage relationships in Orai1 plus STIM1 (a)- and N-FLAG-tagged Orai1 plus STIM1 (b)-cotransfected HEK293 cells and control vector (c)-transfected HEK293 cells. Data represent leak-subtracted current densities (pA/picofarad) evoked by 50-ms voltage ramps from -100 to +100 mV. Right, average CRAC current densities at -80 mV in Orai1 plus STIM1 (n = 11), N-FLAG-tagged Orai1 plus STIM1 (n = 9) and control (n = 9) cells. **, p < 0.01; and ***, p < 0.001.

    Article Snippet: For functional analysis, Orai1 encoding sequence was also subcloned into pCI-neo (Promega, Madison, WI), and mouse STIM1 encoding sequence was subcloned from pApuro-FLAG-tagged mouse STIM1 ( 31 ) into pCI-neo.

    Techniques: Functional Assay, Expressing, Control, Plasmid Preparation, Transfection, Activation Assay

    Predicted topology of Orai1 and a docking model with STIM1. A, schematic diagram of N-FLAG Orai1. Orai1 is a highly glycosylated protein with glycan moiety of ∼8 kDa per subunit. The Orai1 polypeptide has a cytosolic N terminus, four TM segments (yellow columns: S1–S4), and a cytosolic C terminus with a putative coiled-coil sequence (green box) involved in binding with STIM1. The FLAG tag (red box) was introduced in the N terminus. B, illustration of putative docking between Orai1 and STIM1. The three-dimensional reconstruction in this study suggests that the cytoplasmic domain of Orai1 is long enough for direct association with STIM1. STIM1 contains a coiled-coil motif (green box) in the cytoplasm, a single TM segment (yellow column), a sterile α motif domain (purple box), and an EF-hand motif (blue crescent) in the ER lumen.

    Journal:

    Article Title: Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain *

    doi: 10.1074/jbc.M900812200

    Figure Lengend Snippet: Predicted topology of Orai1 and a docking model with STIM1. A, schematic diagram of N-FLAG Orai1. Orai1 is a highly glycosylated protein with glycan moiety of ∼8 kDa per subunit. The Orai1 polypeptide has a cytosolic N terminus, four TM segments (yellow columns: S1–S4), and a cytosolic C terminus with a putative coiled-coil sequence (green box) involved in binding with STIM1. The FLAG tag (red box) was introduced in the N terminus. B, illustration of putative docking between Orai1 and STIM1. The three-dimensional reconstruction in this study suggests that the cytoplasmic domain of Orai1 is long enough for direct association with STIM1. STIM1 contains a coiled-coil motif (green box) in the cytoplasm, a single TM segment (yellow column), a sterile α motif domain (purple box), and an EF-hand motif (blue crescent) in the ER lumen.

    Article Snippet: For functional analysis, Orai1 encoding sequence was also subcloned into pCI-neo (Promega, Madison, WI), and mouse STIM1 encoding sequence was subcloned from pApuro-FLAG-tagged mouse STIM1 ( 31 ) into pCI-neo.

    Techniques: Glycoproteomics, Sequencing, Binding Assay, FLAG-tag, Sterility