snap surface 594  (New England Biolabs)


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  • 93
    Name:
    SNAP Surface 594
    Description:
    SNAP Surface 594 50 nmol
    Catalog Number:
    S9134S
    Price:
    344
    Category:
    Fluorochromes
    Size:
    50 nmol
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    Structured Review

    New England Biolabs snap surface 594
    SNAP Surface 594
    SNAP Surface 594 50 nmol
    https://www.bioz.com/result/snap surface 594/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface 594 - by Bioz Stars, 2021-06
    93/100 stars

    Images

    1) Product Images from "Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins"

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201607095

    TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P
    Figure Legend Snippet: TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P

    Techniques Used: Stable Transfection, Expressing, Transfection, Blocking Assay, Staining, Immunostaining

    2) Product Images from "Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins"

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201607095

    TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P
    Figure Legend Snippet: TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P

    Techniques Used: Stable Transfection, Expressing, Transfection, Blocking Assay, Staining, Immunostaining

    3) Product Images from "Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins"

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201607095

    TULP3/TUB-mediated trafficking to ciliary membrane.  (A) IMCD3 cells stably expressing  SNAP Gpr161 GFP  were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are  > 150 for control and  > 200 for  Tulp3  siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were  > 60 per condition from two independent transfections with  > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing  Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are  > 55 and  > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P
    Figure Legend Snippet: TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P

    Techniques Used: Stable Transfection, Expressing, Transfection, Blocking Assay, Staining, Immunostaining

    Related Articles

    Labeling:

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins
    Article Snippet: Immunofluorescence and microscopy Cells were cultured on coverslips until confluent and starved for indicated periods. .. Surface labeling of starved cells with SNAP–surface block or SNAP–surface 594 at indicated time points in was performed according to the manufacturer’s instructions (New England Biolabs, Inc.). .. In brief, a 4-mM stock solution of SNAP–surface block in DMSO was added to a final concentration of 20 µM in starving medium, and cells were incubated at 37°C and 5% CO2 for 30 min.

    Article Title: Phosphoinositides regulate force-independent interactions between talin, vinculin, and actin
    Article Snippet: Full-length vinculin cell lysates were incubated on Roche cOmplete His-Tag resin for 2 hr at 4°C, then washed with 50 mM Tris-HCl pH 7.8, 500 mM NaCl, 10 mM imidazole, 3 mM β-mercaptoethanol, 1 mM EDTA. .. After washing, proteins were incubated overnight with either 3C protease to remove the SNAP-his tag, or labeled with SNAP-AlexaFluor488 or SNAP-Surface594 (New England Biolabs). .. Following removal or elution from beads, vinculin proteins were then further purified by size-exclusion chromatography using Superdex 200 16/600 column (GE Healthcare) or Superose 6 10/300 column (GE Healthcare) in 50 mM HEPES pH 7.8, 150 mM KCl, 3 mM β-mercaptoethanol, 1 mM EDTA.

    Blocking Assay:

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins
    Article Snippet: Immunofluorescence and microscopy Cells were cultured on coverslips until confluent and starved for indicated periods. .. Surface labeling of starved cells with SNAP–surface block or SNAP–surface 594 at indicated time points in was performed according to the manufacturer’s instructions (New England Biolabs, Inc.). .. In brief, a 4-mM stock solution of SNAP–surface block in DMSO was added to a final concentration of 20 µM in starving medium, and cells were incubated at 37°C and 5% CO2 for 30 min.

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins
    Article Snippet: DSP was from Thermo Fisher Scientific and biotin was from Sigma-Aldrich. .. SNAP–surface block or SNAP–surface 594 was from New England Biolabs, Inc. .. Plasmids pG-LAP1 (pCDNA5/FRT/TO-EGFP-TEV-Stag-X) and pG-LAP5 (pEFα-X-Stag-PreScission-EGFP) were from Addgene ( ).

    Concentration Assay:

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins
    Article Snippet: In brief, a 4-mM stock solution of SNAP–surface block in DMSO was added to a final concentration of 20 µM in starving medium, and cells were incubated at 37°C and 5% CO2 for 30 min. .. Similarly, a 1-mM stock solution of SNAP–surface 594 in DMSO was added to a final concentration of 5 µM in starving medium, and cells were incubated in the dark at 37°C and 5% CO2 for 30 min. ..

    Incubation:

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins
    Article Snippet: In brief, a 4-mM stock solution of SNAP–surface block in DMSO was added to a final concentration of 20 µM in starving medium, and cells were incubated at 37°C and 5% CO2 for 30 min. .. Similarly, a 1-mM stock solution of SNAP–surface 594 in DMSO was added to a final concentration of 5 µM in starving medium, and cells were incubated in the dark at 37°C and 5% CO2 for 30 min. ..

    Article Title: Phosphoinositides regulate force-independent interactions between talin, vinculin, and actin
    Article Snippet: Full-length vinculin cell lysates were incubated on Roche cOmplete His-Tag resin for 2 hr at 4°C, then washed with 50 mM Tris-HCl pH 7.8, 500 mM NaCl, 10 mM imidazole, 3 mM β-mercaptoethanol, 1 mM EDTA. .. After washing, proteins were incubated overnight with either 3C protease to remove the SNAP-his tag, or labeled with SNAP-AlexaFluor488 or SNAP-Surface594 (New England Biolabs). .. Following removal or elution from beads, vinculin proteins were then further purified by size-exclusion chromatography using Superdex 200 16/600 column (GE Healthcare) or Superose 6 10/300 column (GE Healthcare) in 50 mM HEPES pH 7.8, 150 mM KCl, 3 mM β-mercaptoethanol, 1 mM EDTA.

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  • 93
    New England Biolabs snap surface 594
    TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with <t>SNAP–surface</t> block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P
    Snap Surface 594, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/snap surface 594/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    snap surface 594 - by Bioz Stars, 2021-06
    93/100 stars
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    TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P

    Journal: The Journal of Cell Biology

    Article Title: Tubby family proteins are adapters for ciliary trafficking of integral membrane proteins

    doi: 10.1083/jcb.201607095

    Figure Lengend Snippet: TULP3/TUB-mediated trafficking to ciliary membrane. (A) IMCD3 cells stably expressing SNAP Gpr161 GFP were sequentially transfected with 200 nM of the indicated siRNAs twice as indicated in the experimental format on the top left (see Materials and methods). Starved cells were blocked with SNAP–surface block for 30 min, washed, and then were treated with SNAP–surface 594 at indicated time points. Cells were fixed and immunostained for GFP (green) and acetylated tubulin (magenta). Cilia positive and negative for SNAP–surface 594 are marked by white arrows and arrowheads, respectively. The yellow arrowhead points to cilia faintly stained for SNAP–surface 594. Total counted cells are > 150 for control and > 200 for Tulp3 siRNA, respectively. Data represent means ± SD from two or more fields from a single experiment. Bars, 5 µm. (B) NIH 3T3 Flp-In cells stably expressing Gpr161-GFP or Gpr161 V158E -GFP mutants were sequentially transfected with 200 nM of indicated siRNAs twice for 72 h (see Materials and methods) and serum starved for the last 24 h before fixing and immunostaining for GFP, acetylated tubulin, and DNA. Ciliary pixel intensities for GFP are shown. Total counted cells were > 60 per condition from two independent transfections with > 30 cells counted per coverslip. A.U., arbitrary units. Also see Fig. S5 E. (C) IMCD3 Flp-In cells stably and inducibly expressing Myc TULP3 N terminus (1–183 aa) were starved in the presence of doxycycline (Dox) for 20 h (4 µg/ml). After washing, cells were treated ± SAG (500 nM) for indicated time points in starvation medium before fixing and immunostaining for Gpr161, Myc, acetylated tubulin, and DNA. Myc-positive and -negative cells were scored for Gpr161-positive cilia. Total counted cells are > 55 and > 180 for Myc-positive and -negative cells, respectively, for each time point. Data represent means ± SD from three coverslips. Also see Fig. S5 F. (A–C) *, P

    Article Snippet: SNAP–surface block or SNAP–surface 594 was from New England Biolabs, Inc.

    Techniques: Stable Transfection, Expressing, Transfection, Blocking Assay, Staining, Immunostaining