biotinylated protein Search Results


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  • 99
    Vector Laboratories avidin biotin blocking kit
    Avidin Biotin Blocking Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 8750 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher avidin neutravidin biotin binding protein
    Autonomous PIP2 sorting in response to physical structures. a Schematic of the procedure of making PMSD pattern. b SEM images of PDMS micropatterns. c Biotinylated-GPMVs were labeled with BODIPY FL-PIP2, TopFluor-TMR PC, or PE and incubated with PDMS triangular and rectangular patterns coated with <t>NeutrAvidin.</t> Images were taken when GPMV were settled onto a specific pattern. Fold changes, defined as the ratio of fluorescence intensity of a given lipid at the pattern contacting regions over non-contacting regions, were measured for all shapes with or without 10 mM NaN 3 . n ≥ 30, N = 5. Scale bars, 5 µm. *** p
    Avidin Neutravidin Biotin Binding Protein, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend avidin biotin blocking system
    Autonomous PIP2 sorting in response to physical structures. a Schematic of the procedure of making PMSD pattern. b SEM images of PDMS micropatterns. c Biotinylated-GPMVs were labeled with BODIPY FL-PIP2, TopFluor-TMR PC, or PE and incubated with PDMS triangular and rectangular patterns coated with <t>NeutrAvidin.</t> Images were taken when GPMV were settled onto a specific pattern. Fold changes, defined as the ratio of fluorescence intensity of a given lipid at the pattern contacting regions over non-contacting regions, were measured for all shapes with or without 10 mM NaN 3 . n ≥ 30, N = 5. Scale bars, 5 µm. *** p
    Avidin Biotin Blocking System, supplied by BioLegend, used in various techniques. Bioz Stars score: 95/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Biocare Medical avidin biotin kit
    Autonomous PIP2 sorting in response to physical structures. a Schematic of the procedure of making PMSD pattern. b SEM images of PDMS micropatterns. c Biotinylated-GPMVs were labeled with BODIPY FL-PIP2, TopFluor-TMR PC, or PE and incubated with PDMS triangular and rectangular patterns coated with <t>NeutrAvidin.</t> Images were taken when GPMV were settled onto a specific pattern. Fold changes, defined as the ratio of fluorescence intensity of a given lipid at the pattern contacting regions over non-contacting regions, were measured for all shapes with or without 10 mM NaN 3 . n ≥ 30, N = 5. Scale bars, 5 µm. *** p
    Avidin Biotin Kit, supplied by Biocare Medical, used in various techniques. Bioz Stars score: 90/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore avidin biotin complex blotting proteins
    Autonomous PIP2 sorting in response to physical structures. a Schematic of the procedure of making PMSD pattern. b SEM images of PDMS micropatterns. c Biotinylated-GPMVs were labeled with BODIPY FL-PIP2, TopFluor-TMR PC, or PE and incubated with PDMS triangular and rectangular patterns coated with <t>NeutrAvidin.</t> Images were taken when GPMV were settled onto a specific pattern. Fold changes, defined as the ratio of fluorescence intensity of a given lipid at the pattern contacting regions over non-contacting regions, were measured for all shapes with or without 10 mM NaN 3 . n ≥ 30, N = 5. Scale bars, 5 µm. *** p
    Avidin Biotin Complex Blotting Proteins, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher biotinylated
    ILDR2-Fc binds to activated T cells. Binding of ILDR2-hFc was analyzed on resting and activated human CD4 + ( A ) and CD8 + ( B ) T cells. Isolated human CD4 + or CD8 + . Cells were incubated with ILDR2-hFc or isotype control (human IgG1; Synagis) at 100 μg/ml, followed by staining with PE-conjugated anti–human IgG Fc, and evaluated by flow cytometry. The results, shown as line graphs of ILDR2-hFc and isotype control (human IgG1) binding, are representative of two independent experiments. ( C ) Binding kinetics of ILDR2-mFc to resting and activated mouse CD4 + T cells. CD4 + CD25 − . Binding of <t>biotinylated</t> ILDR2-mFc and isotype control (mIgG2a) was tested at different time points of T cell activation and evaluated by flow cytometry. Results are shown as line graphs of ILDR2-mFc and isotype control (mIgG2a) binding.
    Biotinylated, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1230 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Vector Laboratories avidin biotin complex
    ILDR2-Fc binds to activated T cells. Binding of ILDR2-hFc was analyzed on resting and activated human CD4 + ( A ) and CD8 + ( B ) T cells. Isolated human CD4 + or CD8 + . Cells were incubated with ILDR2-hFc or isotype control (human IgG1; Synagis) at 100 μg/ml, followed by staining with PE-conjugated anti–human IgG Fc, and evaluated by flow cytometry. The results, shown as line graphs of ILDR2-hFc and isotype control (human IgG1) binding, are representative of two independent experiments. ( C ) Binding kinetics of ILDR2-mFc to resting and activated mouse CD4 + T cells. CD4 + CD25 − . Binding of <t>biotinylated</t> ILDR2-mFc and isotype control (mIgG2a) was tested at different time points of T cell activation and evaluated by flow cytometry. Results are shown as line graphs of ILDR2-mFc and isotype control (mIgG2a) binding.
    Avidin Biotin Complex, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 9572 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher biotinylated protein a
    Immunoreactivity of recombinant henipavirus nucleocapsid protein and soluble glycoprotein towards different anti-sera from infected and non-infected organisms in a Luminex bead-based assay. Detection of antibodies to recombinant henipavirus N and sG proteins in sera from animals and humans infected naturally (NI) or laboratory infected (LI) with, either HeV, NiV or CedPV, and horses receiving Equivac® HeV vaccine (Vac). Non-infected humans and animals were designated normal. HeV N FL and HeV N CORE together HeV, NiV and CedPV sG were coupled to individual sets of Luminex beads. Binding of specific antibodies were detected using <t>biotinylated</t> <t>Protein</t> A and biotinylated Protein G and streptavidin–phycoerythrin. The results were recorded as the Mean Fluorescent Intensity (MFI) of 100 beads.
    Biotinylated Protein A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Vector Laboratories avidin
    Immunoreactivity of recombinant henipavirus nucleocapsid protein and soluble glycoprotein towards different anti-sera from infected and non-infected organisms in a Luminex bead-based assay. Detection of antibodies to recombinant henipavirus N and sG proteins in sera from animals and humans infected naturally (NI) or laboratory infected (LI) with, either HeV, NiV or CedPV, and horses receiving Equivac® HeV vaccine (Vac). Non-infected humans and animals were designated normal. HeV N FL and HeV N CORE together HeV, NiV and CedPV sG were coupled to individual sets of Luminex beads. Binding of specific antibodies were detected using <t>biotinylated</t> <t>Protein</t> A and biotinylated Protein G and streptavidin–phycoerythrin. The results were recorded as the Mean Fluorescent Intensity (MFI) of 100 beads.
    Avidin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 637 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher neutravidin
    Gene transduction mediated by biotinylated transferrin. Jurkat cells were transduced with BAP II SINDBIS pseudotype either without conjugation, conjugated with <t>neutravidin,</t> or conjugated with neutravidin and biotinylated transferrin. Firefly luciferase was used as a reporter gene, and gene transduction was analysed by measuring the relative luciferase units (RLU) of transduced cells. Representative data of three independent experiments are shown
    Neutravidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher biotinylated antibody
    Noggin does not affect association of LPP with BMP-RII in IVD cells. Cells were incubated with a <t>biotinylated</t> peptide LPPb in the in the absence or presence of noggin (600 ng/ml) and compared with control cells. After a 48-h incubation, cells were washed,
    Biotinylated Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories biotinylated peroxidase
    (A) Structure of the 2-site ELISA for detection of antibodies against the cTfRMAb-ScFv fusion protein. The cTfRMAb-ScFv fusion protein is used as the capture reagent, and the <t>biotinylated</t> cTfRMAb-ScFv fusion protein is used as the detector reagent, along
    Biotinylated Peroxidase, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Cell Signaling Technology Inc biotinylated protein ladder
    fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface <t>biotinylated</t> with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).
    Biotinylated Protein Ladder, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 455 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc biotinylated protein marker
    fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface <t>biotinylated</t> with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).
    Biotinylated Protein Marker, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore biotinylated protein g
    fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface <t>biotinylated</t> with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).
    Biotinylated Protein G, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad biotinylated protein ladder
    fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface <t>biotinylated</t> with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).
    Biotinylated Protein Ladder, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Rockland Immunochemicals biotinylated protein g
    fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface <t>biotinylated</t> with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).
    Biotinylated Protein G, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Autonomous PIP2 sorting in response to physical structures. a Schematic of the procedure of making PMSD pattern. b SEM images of PDMS micropatterns. c Biotinylated-GPMVs were labeled with BODIPY FL-PIP2, TopFluor-TMR PC, or PE and incubated with PDMS triangular and rectangular patterns coated with NeutrAvidin. Images were taken when GPMV were settled onto a specific pattern. Fold changes, defined as the ratio of fluorescence intensity of a given lipid at the pattern contacting regions over non-contacting regions, were measured for all shapes with or without 10 mM NaN 3 . n ≥ 30, N = 5. Scale bars, 5 µm. *** p

    Journal: Nature Communications

    Article Title: A phosphatidylinositol 4,5-bisphosphate redistribution-based sensing mechanism initiates a phagocytosis programing

    doi: 10.1038/s41467-018-06744-7

    Figure Lengend Snippet: Autonomous PIP2 sorting in response to physical structures. a Schematic of the procedure of making PMSD pattern. b SEM images of PDMS micropatterns. c Biotinylated-GPMVs were labeled with BODIPY FL-PIP2, TopFluor-TMR PC, or PE and incubated with PDMS triangular and rectangular patterns coated with NeutrAvidin. Images were taken when GPMV were settled onto a specific pattern. Fold changes, defined as the ratio of fluorescence intensity of a given lipid at the pattern contacting regions over non-contacting regions, were measured for all shapes with or without 10 mM NaN 3 . n ≥ 30, N = 5. Scale bars, 5 µm. *** p

    Article Snippet: To prepare for GPMV/pattern contact experiment, patterns were pre-coated with NeutrAvidin (A2666, Invitrogen) and stored at −20 °C.

    Techniques: Labeling, Incubation, Fluorescence

    ILDR2-Fc binds to activated T cells. Binding of ILDR2-hFc was analyzed on resting and activated human CD4 + ( A ) and CD8 + ( B ) T cells. Isolated human CD4 + or CD8 + . Cells were incubated with ILDR2-hFc or isotype control (human IgG1; Synagis) at 100 μg/ml, followed by staining with PE-conjugated anti–human IgG Fc, and evaluated by flow cytometry. The results, shown as line graphs of ILDR2-hFc and isotype control (human IgG1) binding, are representative of two independent experiments. ( C ) Binding kinetics of ILDR2-mFc to resting and activated mouse CD4 + T cells. CD4 + CD25 − . Binding of biotinylated ILDR2-mFc and isotype control (mIgG2a) was tested at different time points of T cell activation and evaluated by flow cytometry. Results are shown as line graphs of ILDR2-mFc and isotype control (mIgG2a) binding.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: ILDR2 Is a Novel B7-like Protein That Negatively Regulates T Cell Responses

    doi: 10.4049/jimmunol.1700325

    Figure Lengend Snippet: ILDR2-Fc binds to activated T cells. Binding of ILDR2-hFc was analyzed on resting and activated human CD4 + ( A ) and CD8 + ( B ) T cells. Isolated human CD4 + or CD8 + . Cells were incubated with ILDR2-hFc or isotype control (human IgG1; Synagis) at 100 μg/ml, followed by staining with PE-conjugated anti–human IgG Fc, and evaluated by flow cytometry. The results, shown as line graphs of ILDR2-hFc and isotype control (human IgG1) binding, are representative of two independent experiments. ( C ) Binding kinetics of ILDR2-mFc to resting and activated mouse CD4 + T cells. CD4 + CD25 − . Binding of biotinylated ILDR2-mFc and isotype control (mIgG2a) was tested at different time points of T cell activation and evaluated by flow cytometry. Results are shown as line graphs of ILDR2-mFc and isotype control (mIgG2a) binding.

    Article Snippet: For binding of tested proteins, biotinylated (using a DSB-X Biotin Protein Labeling Kit, D-20655; Molecular Probes) ILDR2-mFc or Ig control (mIgG2a) was used.

    Techniques: Binding Assay, Isolation, Incubation, Staining, Flow Cytometry, Cytometry, Activation Assay

    Immunoreactivity of recombinant henipavirus nucleocapsid protein and soluble glycoprotein towards different anti-sera from infected and non-infected organisms in a Luminex bead-based assay. Detection of antibodies to recombinant henipavirus N and sG proteins in sera from animals and humans infected naturally (NI) or laboratory infected (LI) with, either HeV, NiV or CedPV, and horses receiving Equivac® HeV vaccine (Vac). Non-infected humans and animals were designated normal. HeV N FL and HeV N CORE together HeV, NiV and CedPV sG were coupled to individual sets of Luminex beads. Binding of specific antibodies were detected using biotinylated Protein A and biotinylated Protein G and streptavidin–phycoerythrin. The results were recorded as the Mean Fluorescent Intensity (MFI) of 100 beads.

    Journal: Protein Expression and Purification

    Article Title: Structural characterization by transmission electron microscopy and immunoreactivity of recombinant Hendra virus nucleocapsid protein expressed and purified from Escherichia coli

    doi: 10.1016/j.pep.2015.07.008

    Figure Lengend Snippet: Immunoreactivity of recombinant henipavirus nucleocapsid protein and soluble glycoprotein towards different anti-sera from infected and non-infected organisms in a Luminex bead-based assay. Detection of antibodies to recombinant henipavirus N and sG proteins in sera from animals and humans infected naturally (NI) or laboratory infected (LI) with, either HeV, NiV or CedPV, and horses receiving Equivac® HeV vaccine (Vac). Non-infected humans and animals were designated normal. HeV N FL and HeV N CORE together HeV, NiV and CedPV sG were coupled to individual sets of Luminex beads. Binding of specific antibodies were detected using biotinylated Protein A and biotinylated Protein G and streptavidin–phycoerythrin. The results were recorded as the Mean Fluorescent Intensity (MFI) of 100 beads.

    Article Snippet: The bound antibodies were then detected using biotinylated Protein A (Pierce) and biotinylated Protein G (Pierce) and streptavidin–phycoerythrin (Qiagen).

    Techniques: Recombinant, Infection, Luminex, Bead-based Assay, Binding Assay

    Close proximity of cadherin molecules in a cis orientation cooperatively increases the probability of trans cadherin binding. ( A ) Schematic of AFM tip and substrate functionalized with cadherin–Fc dimers for single-molecule force measurements. The tip and surface were functionalized with PEG linkers, some of which were decorated with streptavidin molecules. Cadherin–Fc dimers were immobilized on biotinylated protein G attached to the streptavidin molecules on the tip and surface. The flexible PEG linkers enable unhindered interactions between opposing cadherins during tip–substrate encounters. Cadherin–Fc dimers were immobilized on the AFM tip and substrate at a surface density of 34 ± 16 dimers per μm 2 . ( B ) Schematic of AFM tip and substrate functionalized with biotinylated cadherin monomers for single-molecule force measurements. The biotinylated cadherin monomers were immobilized on an AFM tip and substrate via streptavidin-PEG tethers at a surface densities of 65 ± 18 monomers per μm 2 ; thus, the protein surface density for the cadherin monomers and cadherin–Fc dimer constructs are similar. ( C ) A typical force curve showing the unbinding of a single cadherin molecule. The tip and the substrate decorated with cadherins were bought into contact for either 1,100, 340, or 115 ms so that cadherins on the tip and substrate formed an adhesive complex. When the tip was withdrawn from the substrate a force was exerted, and above a critical force the adhesive complex ruptured. Forces between the cadherin–Fc dimers were measured 7,995 times in 2.5 mM Ca 2+ , yielding 509 binding events, and 5,931 times in 2 mM EGTA, yielding 123 binding events. Forces between the cadherin monomers were measured 5,949 times in 2.5 mM Ca 2+ , yielding 113 binding events, and 5,846 times in 2 mM EGTA, yielding 28 binding events. ( D ) Histogram of cadherin–Fc dimer binding events measured in 2.5 mM Ca 2+ (solid gray bars) and 2 mM EGTA (hatched green bars). The binding events measured in 2.5 mM Ca 2+ were fitted to a Gaussian distribution with a peak force of 53 ± 27 pN. The binding probability of cadherin–Fc dimers at contact times of 1,100, 340, or 115 ms was 8.6%, 6.6%, and 3.7%, respectively. The histogram includes the forces measured at all 3 tip-surface contact times. ( E ) Histogram of cadherin monomer binding events measured in 2.5 mM Ca 2+ (solid gray bars) and 2 mM EGTA (hatched green bars). The histogram includes the forces measured at all 3 tip-surface contact times (see above). The binding events measured in 2.5 mM Ca 2+ were fitted to a Gaussian distribution with a peak force of 64 ± 27 pN. Although the bond strengths of the monomer and cadherin–Fc dimer adhesive complexes are similar, the cadherin–Fc dimer has a higher probability of binding than a cadherin monomer. The total probability of cadherin monomer binding at contact times of 1,100, 340, or 115 ms was 2.5%, 2.2%, and 1.1% respectively. Thus, the cadherin–Fc dimer binding probability was 3.5, 3.1, and 3.6 times higher than the probability of monomer interactions at these tip-surface contact times.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Resolving cadherin interactions and binding cooperativity at the single-molecule level

    doi: 10.1073/pnas.0811350106

    Figure Lengend Snippet: Close proximity of cadherin molecules in a cis orientation cooperatively increases the probability of trans cadherin binding. ( A ) Schematic of AFM tip and substrate functionalized with cadherin–Fc dimers for single-molecule force measurements. The tip and surface were functionalized with PEG linkers, some of which were decorated with streptavidin molecules. Cadherin–Fc dimers were immobilized on biotinylated protein G attached to the streptavidin molecules on the tip and surface. The flexible PEG linkers enable unhindered interactions between opposing cadherins during tip–substrate encounters. Cadherin–Fc dimers were immobilized on the AFM tip and substrate at a surface density of 34 ± 16 dimers per μm 2 . ( B ) Schematic of AFM tip and substrate functionalized with biotinylated cadherin monomers for single-molecule force measurements. The biotinylated cadherin monomers were immobilized on an AFM tip and substrate via streptavidin-PEG tethers at a surface densities of 65 ± 18 monomers per μm 2 ; thus, the protein surface density for the cadherin monomers and cadherin–Fc dimer constructs are similar. ( C ) A typical force curve showing the unbinding of a single cadherin molecule. The tip and the substrate decorated with cadherins were bought into contact for either 1,100, 340, or 115 ms so that cadherins on the tip and substrate formed an adhesive complex. When the tip was withdrawn from the substrate a force was exerted, and above a critical force the adhesive complex ruptured. Forces between the cadherin–Fc dimers were measured 7,995 times in 2.5 mM Ca 2+ , yielding 509 binding events, and 5,931 times in 2 mM EGTA, yielding 123 binding events. Forces between the cadherin monomers were measured 5,949 times in 2.5 mM Ca 2+ , yielding 113 binding events, and 5,846 times in 2 mM EGTA, yielding 28 binding events. ( D ) Histogram of cadherin–Fc dimer binding events measured in 2.5 mM Ca 2+ (solid gray bars) and 2 mM EGTA (hatched green bars). The binding events measured in 2.5 mM Ca 2+ were fitted to a Gaussian distribution with a peak force of 53 ± 27 pN. The binding probability of cadherin–Fc dimers at contact times of 1,100, 340, or 115 ms was 8.6%, 6.6%, and 3.7%, respectively. The histogram includes the forces measured at all 3 tip-surface contact times. ( E ) Histogram of cadherin monomer binding events measured in 2.5 mM Ca 2+ (solid gray bars) and 2 mM EGTA (hatched green bars). The histogram includes the forces measured at all 3 tip-surface contact times (see above). The binding events measured in 2.5 mM Ca 2+ were fitted to a Gaussian distribution with a peak force of 64 ± 27 pN. Although the bond strengths of the monomer and cadherin–Fc dimer adhesive complexes are similar, the cadherin–Fc dimer has a higher probability of binding than a cadherin monomer. The total probability of cadherin monomer binding at contact times of 1,100, 340, or 115 ms was 2.5%, 2.2%, and 1.1% respectively. Thus, the cadherin–Fc dimer binding probability was 3.5, 3.1, and 3.6 times higher than the probability of monomer interactions at these tip-surface contact times.

    Article Snippet: To immobilize Fc–cadherin dimers, the streptavidins were decorated with biotinylated protein G (Pierce Biotechnology), and the Fc region of the cadherin dimer was specifically bound to the protein G. Fluorescently-labeled cadherin molecules were observed by using a home-built prism-type total internal reflection fluorescence microscope.

    Techniques: Binding Assay, Construct, Mass Spectrometry

    Fig. 4. ( A ) Western blot detection of cell surface-bound E2. Biotinylated HepG2 cells were incubated in the presence (lanes 1 and 3) or in the absence (lanes 2 and 4) of E2 recombinant protein. Bound E2 was cross-linked with DTSSP and the E2–receptor complexes were immunoprecipitated with an antibody against the His tag of the E2 recombinant protein. Samples eluted under both non-reducing (lanes 1 and 2) and reducing (lanes 3 and 4) conditions were loaded onto a 10% SDS–PAGE gel. E2 protein was detected as a monomer under reducing conditions (lane 3) and at a higher molecular weight under non- reducing conditions (lane 1), by using an anti-E2 rat mAb followed by anti-rat HRP-conjugated. ( B ) Immunoblot detection of biotinylated cell surface proteins interacting with E2. The reactivity with HRP conjugated streptavidin revealed a biotinylated protein band at 82 kDa only under reducing conditions (lane 3).

    Journal: The EMBO Journal

    Article Title: The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus

    doi: 10.1093/emboj/cdf529

    Figure Lengend Snippet: Fig. 4. ( A ) Western blot detection of cell surface-bound E2. Biotinylated HepG2 cells were incubated in the presence (lanes 1 and 3) or in the absence (lanes 2 and 4) of E2 recombinant protein. Bound E2 was cross-linked with DTSSP and the E2–receptor complexes were immunoprecipitated with an antibody against the His tag of the E2 recombinant protein. Samples eluted under both non-reducing (lanes 1 and 2) and reducing (lanes 3 and 4) conditions were loaded onto a 10% SDS–PAGE gel. E2 protein was detected as a monomer under reducing conditions (lane 3) and at a higher molecular weight under non- reducing conditions (lane 1), by using an anti-E2 rat mAb followed by anti-rat HRP-conjugated. ( B ) Immunoblot detection of biotinylated cell surface proteins interacting with E2. The reactivity with HRP conjugated streptavidin revealed a biotinylated protein band at 82 kDa only under reducing conditions (lane 3).

    Article Snippet: Samples obtained by boiling the beads were loaded on to an SDS–PAGE gel and analyzed by western blot for detection of biotinylated proteins using horseradish peroxidase (HRP)-conjugated streptavidin (Pierce) diluted 1:25 000 in Tris-buffered saline 0.05% Tween 20 (TBST), 2% bovine serum albumin (BSA).

    Techniques: Western Blot, Incubation, Recombinant, Immunoprecipitation, SDS Page, Molecular Weight

    Gene transduction mediated by biotinylated transferrin. Jurkat cells were transduced with BAP II SINDBIS pseudotype either without conjugation, conjugated with neutravidin, or conjugated with neutravidin and biotinylated transferrin. Firefly luciferase was used as a reporter gene, and gene transduction was analysed by measuring the relative luciferase units (RLU) of transduced cells. Representative data of three independent experiments are shown

    Journal: The journal of gene medicine

    Article Title: A versatile targeting system with lentiviral vectors bearing the biotin-adaptor peptide

    doi: 10.1002/jgm.1345

    Figure Lengend Snippet: Gene transduction mediated by biotinylated transferrin. Jurkat cells were transduced with BAP II SINDBIS pseudotype either without conjugation, conjugated with neutravidin, or conjugated with neutravidin and biotinylated transferrin. Firefly luciferase was used as a reporter gene, and gene transduction was analysed by measuring the relative luciferase units (RLU) of transduced cells. Representative data of three independent experiments are shown

    Article Snippet: Neutravidin was purchased from Pierce (Rockford, IL, USA).

    Techniques: Transduction, Conjugation Assay, Luciferase

    ]. 2.2 1L1L has two flexible linkers (Gly-Gly-Gly-Gly-Ser) and AVR II-Bst E II cloning sites at amino acid 71 of the E2 protein. BAP Sindbis contains a biotin acceptor peptide (BAP) derived from Escherichia coli biotin holoenzyme synthetase between the two flexible linkers. BBAPH contains BAP at the first amino acid of the E2 protein, and BAP II SINDBIS contains two BAP at both positions of the E2 protein. (b) The schematic strategy to conjugate virus with anti-human or rat transferrin receptor 1 and transferrin. BAP II SINDBIS envelope proteins were covalently conjugated with biotin. Anti-human or rat transferrin receptor antibodies were fused with avidin, designated anti-huTfR IgG-Av or anti-ratTfR IgG3-Av, respectively. Anti-huTfR IgG-Av or anti-ratTfR IgG3-Av can be conjugated with the BAP II SINDBIS envelope protein through the interaction of avidin and the biotin of the envelope protein. Neutravidin has four biotin binding sites. Thus, one neutravidin can bind both the biotinylated BAP II SINDBIS envelope protein and biotinlynated transferrin, which results in bridging the pseudotyped virus with transferrin

    Journal: The journal of gene medicine

    Article Title: A versatile targeting system with lentiviral vectors bearing the biotin-adaptor peptide

    doi: 10.1002/jgm.1345

    Figure Lengend Snippet: ]. 2.2 1L1L has two flexible linkers (Gly-Gly-Gly-Gly-Ser) and AVR II-Bst E II cloning sites at amino acid 71 of the E2 protein. BAP Sindbis contains a biotin acceptor peptide (BAP) derived from Escherichia coli biotin holoenzyme synthetase between the two flexible linkers. BBAPH contains BAP at the first amino acid of the E2 protein, and BAP II SINDBIS contains two BAP at both positions of the E2 protein. (b) The schematic strategy to conjugate virus with anti-human or rat transferrin receptor 1 and transferrin. BAP II SINDBIS envelope proteins were covalently conjugated with biotin. Anti-human or rat transferrin receptor antibodies were fused with avidin, designated anti-huTfR IgG-Av or anti-ratTfR IgG3-Av, respectively. Anti-huTfR IgG-Av or anti-ratTfR IgG3-Av can be conjugated with the BAP II SINDBIS envelope protein through the interaction of avidin and the biotin of the envelope protein. Neutravidin has four biotin binding sites. Thus, one neutravidin can bind both the biotinylated BAP II SINDBIS envelope protein and biotinlynated transferrin, which results in bridging the pseudotyped virus with transferrin

    Article Snippet: Neutravidin was purchased from Pierce (Rockford, IL, USA).

    Techniques: Clone Assay, Derivative Assay, Avidin-Biotin Assay, Binding Assay

    Noggin does not affect association of LPP with BMP-RII in IVD cells. Cells were incubated with a biotinylated peptide LPPb in the in the absence or presence of noggin (600 ng/ml) and compared with control cells. After a 48-h incubation, cells were washed,

    Journal: The Journal of Biological Chemistry

    Article Title: Link Protein N-terminal Peptide Binds to Bone Morphogenetic Protein (BMP) Type II Receptor and Drives Matrix Protein Expression in Rabbit Intervertebral Disc Cells *

    doi: 10.1074/jbc.M113.451948

    Figure Lengend Snippet: Noggin does not affect association of LPP with BMP-RII in IVD cells. Cells were incubated with a biotinylated peptide LPPb in the in the absence or presence of noggin (600 ng/ml) and compared with control cells. After a 48-h incubation, cells were washed,

    Article Snippet: To capture biotinylated LPP-bound protein, the supernatant containing biotinylated peptide-protein conjugates was passed through a monomeric avidin-Sepharose column (Pierce).

    Techniques: Incubation

    LPP directly binds to BMP-RII in IVD cells. Cells were incubated with a biotinylated peptide LPPb or a biotinylated control peptide LPRb and compared with untreated cells ( Ctrl ). A , after a 48-h incubation, cells were labeled with Alexa Fluor 568-conjugated

    Journal: The Journal of Biological Chemistry

    Article Title: Link Protein N-terminal Peptide Binds to Bone Morphogenetic Protein (BMP) Type II Receptor and Drives Matrix Protein Expression in Rabbit Intervertebral Disc Cells *

    doi: 10.1074/jbc.M113.451948

    Figure Lengend Snippet: LPP directly binds to BMP-RII in IVD cells. Cells were incubated with a biotinylated peptide LPPb or a biotinylated control peptide LPRb and compared with untreated cells ( Ctrl ). A , after a 48-h incubation, cells were labeled with Alexa Fluor 568-conjugated

    Article Snippet: To capture biotinylated LPP-bound protein, the supernatant containing biotinylated peptide-protein conjugates was passed through a monomeric avidin-Sepharose column (Pierce).

    Techniques: Incubation, Labeling

    (A) Structure of the 2-site ELISA for detection of antibodies against the cTfRMAb-ScFv fusion protein. The cTfRMAb-ScFv fusion protein is used as the capture reagent, and the biotinylated cTfRMAb-ScFv fusion protein is used as the detector reagent, along

    Journal: Molecular pharmaceutics

    Article Title: Receptor-Mediated Abeta Amyloid Antibody Targeting to Alzheimer's Disease Mouse Brain

    doi: 10.1021/mp1003515

    Figure Lengend Snippet: (A) Structure of the 2-site ELISA for detection of antibodies against the cTfRMAb-ScFv fusion protein. The cTfRMAb-ScFv fusion protein is used as the capture reagent, and the biotinylated cTfRMAb-ScFv fusion protein is used as the detector reagent, along

    Article Snippet: The biotinylation of the cTfRMAb-ScFv fusion protein was confirmed by SDS-PAGE and Western blotting, where the blot was probed with avidin and biotinylated peroxidase.

    Techniques: Enzyme-linked Immunosorbent Assay

    fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface biotinylated with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).

    Journal: The Journal of Biological Chemistry

    Article Title: Functional Expression of the Na-K-2Cl Cotransporter NKCC2 in Mammalian Cells Fails to Confirm the Dominant-negative Effect of the AF Splice Variant *

    doi: 10.1074/jbc.M109.060004

    Figure Lengend Snippet: fNKCC2AF: glycosylation, surface expression, and 86 Rb uptake. A , cell lysates from HEK-293 cells transfected with fNKCC2AF were incubated with (+) or without (−) PNGase F, subjected to SDS-PAGE, immunoblotted with anti-NKCC2 ( left panel ), and re-probed with N1 antibody ( right panel ). Molecular size markers (in kDa) and cross-reaction of anti-NKCC2 with NKCC1 (*) are indicated on the left and NKCC2AF is marked by an arrow on the right. B , 48 h after transfection with fNKCC2AF HEK-293 cells were surface biotinylated with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, run on SDS-PAGE, immunoblotted with anti-NKCC2 ( left ), and re-probed with N1 to detect endogenous NKCC1 ( right ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins ( S ) compared with total proteins ( T ). C , untransfected ( HEK ) and fNKCC2AF-transfected ( AF ) HEK-293 cells, and D , stable fNKCC2A ( A ) and stable fNKCC2A cells co-expressing fNKCC2AF ( A+AF ) were grown to confluence (2–3 days). 86 Rb uptake was assessed following incubation of cells in isotonic (basal conditions, open bars ) or hypotonic/low-chloride ( filled bars ) medium. Bumetanide-sensitive 86 Rb uptakes are shown as mean ± S.E. Expression of fNKCC2AF has no significant ( n.s. ) effect on 86 Rb uptakes by HEK-293 cells ( C ) ( n ≥ 4) or HEK-293 cells stably expressing fNKCC2A ( D ) ( n ≥ 3).

    Article Snippet: Biotinylated protein ladder (Cell Signaling Technology, Danvers, MA) and MagicMark XP marker (Invitrogen) were used for estimation of molecular weights.

    Techniques: Expressing, Transfection, Incubation, SDS Page, Stable Transfection

    fNKCC2 glycosylation and appearance in the cell surface. A , whole cell lysates from HEK-293 cells transfected with fNKCC2A, -B, and -F were treated with (+) or without (−) PNGase F, subjected to SDS-PAGE, and immunoblotted with anti-NKCC2 ( left panel ) and re-probed with T4 ( right panel ). Exposure time was increased to detect fNKCC2F (shown by vertical line ). B , detection of surface proteins. 48 h after transfection HEK-293 cells were surface biotinylated with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, eluted, run on SDS-PAGE, and immunoblotted with anti-NKCC2 ( left panels ) and re-probed with N1 to detect endogenous NKCC1 ( right panels ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins compared with total proteins. Molecular mass markers (in kDa) are indicated.

    Journal: The Journal of Biological Chemistry

    Article Title: Functional Expression of the Na-K-2Cl Cotransporter NKCC2 in Mammalian Cells Fails to Confirm the Dominant-negative Effect of the AF Splice Variant *

    doi: 10.1074/jbc.M109.060004

    Figure Lengend Snippet: fNKCC2 glycosylation and appearance in the cell surface. A , whole cell lysates from HEK-293 cells transfected with fNKCC2A, -B, and -F were treated with (+) or without (−) PNGase F, subjected to SDS-PAGE, and immunoblotted with anti-NKCC2 ( left panel ) and re-probed with T4 ( right panel ). Exposure time was increased to detect fNKCC2F (shown by vertical line ). B , detection of surface proteins. 48 h after transfection HEK-293 cells were surface biotinylated with Sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated from cell lysates with streptavidin beads, eluted, run on SDS-PAGE, and immunoblotted with anti-NKCC2 ( left panels ) and re-probed with N1 to detect endogenous NKCC1 ( right panels ). Twice as much protein was loaded and longer exposure times were used to detect surface proteins compared with total proteins. Molecular mass markers (in kDa) are indicated.

    Article Snippet: Biotinylated protein ladder (Cell Signaling Technology, Danvers, MA) and MagicMark XP marker (Invitrogen) were used for estimation of molecular weights.

    Techniques: Transfection, SDS Page