anti gfp roche cat 11814460001 rrid ab 390913  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti gfp roche cat 11814460001 rrid ab 390913
    Anti Gfp Roche Cat 11814460001 Rrid Ab 390913, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse anti gfp antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc mouse anti gfp antibody
    Mouse Anti Gfp Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    antibody anti mad1 anti gfp or anti myc  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc antibody anti mad1 anti gfp or anti myc
    Antibody Anti Mad1 Anti Gfp Or Anti Myc, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    gfp antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gfp antibody
    (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions <t>containing</t> <t>EphA2-GFP</t> were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.
    Gfp Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Cholesterol inhibits assembly and activation of the EphA2 receptor"

    Article Title: Cholesterol inhibits assembly and activation of the EphA2 receptor

    Journal: bioRxiv

    doi: 10.1101/2024.06.10.598255

    (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions containing EphA2-GFP were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.
    Figure Legend Snippet: (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions containing EphA2-GFP were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.

    Techniques Used: Sample Prep, Membrane, Microscopy, Imaging

    (A) Schematic of DIBMALP containing an EphA2-GFP monomer. (B) Step distribution of control DIBMALPs (black) or those formed from cells treated with EA1 (pink), MβCD (blue) or both (magenta). (C) Oligomeric distribution calculated from data in panel B. (D) Schematic representing DDM micelles containing EphA2-GFP. (E) Step distribution of DDM-solubilized EphA2-GFP in the same conditions as in DIBMALPs. (F) Oligomeric distribution of DDM-solubilized EphA2-GFP photobleaching data. p -values are from two-way ANOVA followed by Tukey multiple comparison test.*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.
    Figure Legend Snippet: (A) Schematic of DIBMALP containing an EphA2-GFP monomer. (B) Step distribution of control DIBMALPs (black) or those formed from cells treated with EA1 (pink), MβCD (blue) or both (magenta). (C) Oligomeric distribution calculated from data in panel B. (D) Schematic representing DDM micelles containing EphA2-GFP. (E) Step distribution of DDM-solubilized EphA2-GFP in the same conditions as in DIBMALPs. (F) Oligomeric distribution of DDM-solubilized EphA2-GFP photobleaching data. p -values are from two-way ANOVA followed by Tukey multiple comparison test.*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

    Techniques Used: Comparison

    antibodies rabbit anti stim1 mab d88e10 cell signaling 5668 chicken anti gfp abcam ab13970  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc antibodies rabbit anti stim1 mab d88e10 cell signaling 5668 chicken anti gfp abcam ab13970
    Antibodies Rabbit Anti Stim1 Mab D88e10 Cell Signaling 5668 Chicken Anti Gfp Abcam Ab13970, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    antibody anti mad1 anti gfp or anti myc  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc antibody anti mad1 anti gfp or anti myc
    (A) An alphafold model of <t>Mad1-Mad2</t> hetero-tetramer indicating conserved domains in CnMad1: the Mad2 interaction site; 2 putative Bub1 binding motifs (RLK) and the structurally conserved C-terminal domain (RWD). (B) The mad1 Δ strain is benomyl sensitive and can be complemented with an ectopic <t>GFP-Mad1</t> construct. Strains were grown on YPD plates containing the indicated concentrations of benomyl for 3 days at 30°C. (C) Anti-Mad1 immunoblot confirms the mad1 Δ strain. The three strains indicated were grown to log phase, harvested and whole cell extracts generated. The upper band (~115kD, labelled *) recognised by the anti-Mad1 antibody is a cross-reacting band and is used here as a loading control. (D) Benomyl sensitivity of the mad2 Δ and complementation with ectopic GALp-MAD2 . Strains were grown on plates (with 2% glucose or 2% galactose) containing the indicated concentrations of benomyl for 3 days at 30°C. (E) Plate reader experiments confirm that mad1 , mad2 and mps1 mutants are all temperature sensitive, with populations expanding slower than wild-type. Cells were diluted to OD 600 0.2 and grown with shaking at the indicated temperatures. For viability assay (colony forming units) see . (F) The mad1 , mad2 double mutant does not display a synthetic phenotype. mad1 , mad2 , mad1 , mad2 double and mps1 strains were diluted, plated and grown on YPD plates at the indicated temperatures and drug concentrations for 3 days.
    Antibody Anti Mad1 Anti Gfp Or Anti Myc, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Conserved signalling functions for Mps1, Mad1 and Mad2 in the Cryptococcus neoformans spindle checkpoint"

    Article Title: Conserved signalling functions for Mps1, Mad1 and Mad2 in the Cryptococcus neoformans spindle checkpoint

    Journal: PLOS Genetics

    doi: 10.1371/journal.pgen.1011302

    (A) An alphafold model of Mad1-Mad2 hetero-tetramer indicating conserved domains in CnMad1: the Mad2 interaction site; 2 putative Bub1 binding motifs (RLK) and the structurally conserved C-terminal domain (RWD). (B) The mad1 Δ strain is benomyl sensitive and can be complemented with an ectopic GFP-Mad1 construct. Strains were grown on YPD plates containing the indicated concentrations of benomyl for 3 days at 30°C. (C) Anti-Mad1 immunoblot confirms the mad1 Δ strain. The three strains indicated were grown to log phase, harvested and whole cell extracts generated. The upper band (~115kD, labelled *) recognised by the anti-Mad1 antibody is a cross-reacting band and is used here as a loading control. (D) Benomyl sensitivity of the mad2 Δ and complementation with ectopic GALp-MAD2 . Strains were grown on plates (with 2% glucose or 2% galactose) containing the indicated concentrations of benomyl for 3 days at 30°C. (E) Plate reader experiments confirm that mad1 , mad2 and mps1 mutants are all temperature sensitive, with populations expanding slower than wild-type. Cells were diluted to OD 600 0.2 and grown with shaking at the indicated temperatures. For viability assay (colony forming units) see . (F) The mad1 , mad2 double mutant does not display a synthetic phenotype. mad1 , mad2 , mad1 , mad2 double and mps1 strains were diluted, plated and grown on YPD plates at the indicated temperatures and drug concentrations for 3 days.
    Figure Legend Snippet: (A) An alphafold model of Mad1-Mad2 hetero-tetramer indicating conserved domains in CnMad1: the Mad2 interaction site; 2 putative Bub1 binding motifs (RLK) and the structurally conserved C-terminal domain (RWD). (B) The mad1 Δ strain is benomyl sensitive and can be complemented with an ectopic GFP-Mad1 construct. Strains were grown on YPD plates containing the indicated concentrations of benomyl for 3 days at 30°C. (C) Anti-Mad1 immunoblot confirms the mad1 Δ strain. The three strains indicated were grown to log phase, harvested and whole cell extracts generated. The upper band (~115kD, labelled *) recognised by the anti-Mad1 antibody is a cross-reacting band and is used here as a loading control. (D) Benomyl sensitivity of the mad2 Δ and complementation with ectopic GALp-MAD2 . Strains were grown on plates (with 2% glucose or 2% galactose) containing the indicated concentrations of benomyl for 3 days at 30°C. (E) Plate reader experiments confirm that mad1 , mad2 and mps1 mutants are all temperature sensitive, with populations expanding slower than wild-type. Cells were diluted to OD 600 0.2 and grown with shaking at the indicated temperatures. For viability assay (colony forming units) see . (F) The mad1 , mad2 double mutant does not display a synthetic phenotype. mad1 , mad2 , mad1 , mad2 double and mps1 strains were diluted, plated and grown on YPD plates at the indicated temperatures and drug concentrations for 3 days.

    Techniques Used: Binding Assay, Construct, Western Blot, Generated, Viability Assay, Mutagenesis

    (A) mad1 , mad2 and mps1 strains fail to maintain a large-budded nocodazole arrest. The strains indicated were grown to log phase and then nocodazole was added to a final concentration of 2.5 μg/ml in YPD media for 3 hours. Scale bar is 10μm. (B) Quantitation of the large-budded cells (bud diameter >4μm) observed at the 3 hour time point during this nocodazole challenge. This experiment was repeated 3 times and 300 cells were counted for each strain per experiment. (C) Images taken from a microfluidics analysis of this behaviour in nocodazole (for H99, mad1 , mad2 and mps1 mutants). Cells were pre-grown in synthetic complete media supplemented with 0.2g/ml glucose and 0.05% w/v bovine serum albumin (Sigma) and then injected into the microfluidics device and analysed for 4 hours (+/- 2.5μg/ml nocodazole). Scale bar is 10μm. (D) Quantitation of the microfluidics experiment: movies were analysed manually for re-budding. This experiment was repeated three times and at least 100 cells counted per strain each time.
    Figure Legend Snippet: (A) mad1 , mad2 and mps1 strains fail to maintain a large-budded nocodazole arrest. The strains indicated were grown to log phase and then nocodazole was added to a final concentration of 2.5 μg/ml in YPD media for 3 hours. Scale bar is 10μm. (B) Quantitation of the large-budded cells (bud diameter >4μm) observed at the 3 hour time point during this nocodazole challenge. This experiment was repeated 3 times and 300 cells were counted for each strain per experiment. (C) Images taken from a microfluidics analysis of this behaviour in nocodazole (for H99, mad1 , mad2 and mps1 mutants). Cells were pre-grown in synthetic complete media supplemented with 0.2g/ml glucose and 0.05% w/v bovine serum albumin (Sigma) and then injected into the microfluidics device and analysed for 4 hours (+/- 2.5μg/ml nocodazole). Scale bar is 10μm. (D) Quantitation of the microfluidics experiment: movies were analysed manually for re-budding. This experiment was repeated three times and at least 100 cells counted per strain each time.

    Techniques Used: Concentration Assay, Quantitation Assay, Injection

    (A) Strains expressing GFP-Mad1 were stained with Hoechst to label their DNA in cultures grown with and without the addition of 2.5μg/ml nocodazole. Scale bar is 10μm. (B) Five representative stages of mitosis are shown here, from live imaging of a strain expressing both GFP-Mad1 and mCherry-Tub4 (the gamma tubulin construct labels the spindle poles in mitosis). Scale bar is 10μm. (C) In nocodazole arrested cells, GFP-Mad1 is close to but does not co-localise with spindle poles (mCherry-Tub4). Scale bar is 10μm. (D) In nocodazole arrested cells, GFP-Mad1 does co-localise with the centromere marker Cse4-mCherry. Scale bar is 10μm. (E) In interphase, cycling cells, GFP-Mad1 rarely co-localises with Cse4-mCherry. Scale is 10μm.
    Figure Legend Snippet: (A) Strains expressing GFP-Mad1 were stained with Hoechst to label their DNA in cultures grown with and without the addition of 2.5μg/ml nocodazole. Scale bar is 10μm. (B) Five representative stages of mitosis are shown here, from live imaging of a strain expressing both GFP-Mad1 and mCherry-Tub4 (the gamma tubulin construct labels the spindle poles in mitosis). Scale bar is 10μm. (C) In nocodazole arrested cells, GFP-Mad1 is close to but does not co-localise with spindle poles (mCherry-Tub4). Scale bar is 10μm. (D) In nocodazole arrested cells, GFP-Mad1 does co-localise with the centromere marker Cse4-mCherry. Scale bar is 10μm. (E) In interphase, cycling cells, GFP-Mad1 rarely co-localises with Cse4-mCherry. Scale is 10μm.

    Techniques Used: Expressing, Staining, Imaging, Construct, Marker

    GFP-Mad1 was immunoprecipitated from extracts using GFP-TRAP and immunoprecipitates were run into an SDS-PAGE gel, cut out and digested into peptides with trypsin before analysis on a Orbitrap Fusion Lumos Tribrid mass spectrometer. (A) Analysis of GFP-Mad1 interactors: untagged, nocodazole-arrested cells v. nocodazole-arrested GFP-Mad1. This volcano plot shows both the difference (mean, label free quantitation LFQ difference, on the x-axis) and their statistical confidence (-log 10 P-value of Perseus statistical test, on the y-axis) between polypeptides identified in the GFP-Mad1 and untagged control pull-downs (n = 3 for each purification). The -log 10 P-value of 1.3 used here corresponds to a p-value cutoff of 0.05. (B) Analysis of mitotic GFP-Mad1 interactors: cycling GFP-Mad1 cells v. nocodazole-arrested GFP-Mad1. (C) Alignment of the Mad-RLK region in C . neoformans , Homo sapiens , Schizosaccharomyces pombe and Saccharomyces cerevisiae . Only the second RLK (residues 567–9) is within a conserved region. (D) GFP-Mad1 and mCherry-Bub1 co-localise in mitotic cells. Scale bar is 10μm. (E) Co-immunoprecipitation of Mad1 and Bub1 is Mad1-RLK-dependent. mCherry-Bub1 was immunoprecipitated from the five strains indicated. Bub1 immunoprecipitates were washed, split into two sets, separated by SDS-PAGE, then immunoblotted with anti-GFP and anti-Mad1 antibodies.
    Figure Legend Snippet: GFP-Mad1 was immunoprecipitated from extracts using GFP-TRAP and immunoprecipitates were run into an SDS-PAGE gel, cut out and digested into peptides with trypsin before analysis on a Orbitrap Fusion Lumos Tribrid mass spectrometer. (A) Analysis of GFP-Mad1 interactors: untagged, nocodazole-arrested cells v. nocodazole-arrested GFP-Mad1. This volcano plot shows both the difference (mean, label free quantitation LFQ difference, on the x-axis) and their statistical confidence (-log 10 P-value of Perseus statistical test, on the y-axis) between polypeptides identified in the GFP-Mad1 and untagged control pull-downs (n = 3 for each purification). The -log 10 P-value of 1.3 used here corresponds to a p-value cutoff of 0.05. (B) Analysis of mitotic GFP-Mad1 interactors: cycling GFP-Mad1 cells v. nocodazole-arrested GFP-Mad1. (C) Alignment of the Mad-RLK region in C . neoformans , Homo sapiens , Schizosaccharomyces pombe and Saccharomyces cerevisiae . Only the second RLK (residues 567–9) is within a conserved region. (D) GFP-Mad1 and mCherry-Bub1 co-localise in mitotic cells. Scale bar is 10μm. (E) Co-immunoprecipitation of Mad1 and Bub1 is Mad1-RLK-dependent. mCherry-Bub1 was immunoprecipitated from the five strains indicated. Bub1 immunoprecipitates were washed, split into two sets, separated by SDS-PAGE, then immunoblotted with anti-GFP and anti-Mad1 antibodies.

    Techniques Used: Immunoprecipitation, SDS Page, Mass Spectrometry, Quantitation Assay, Purification

    (A) P GAL7 -MPS1 was induced for 5 hours with 2% Galactose, or 2% glucose as the un-induced control. Scale bar is 10μm. (B) Quantitation of this mitotic arrest. Short spindles were scored every hour in at least 100 cells per condition at each time point. This experiment was repeated 3 times and displayed here as the mean +/- confidence levels. (C) Immunoblot of Mps1 levels at each hourly time point after Gal induction. Whole cell extracts were prepared by bead-beating, separated by SDS-PAGE and immunoblotted with the 9E10 anti-myc monoclonal antibody. * indicates a cross-reacting band, used here as a loading control. (D) P GAL7 -MPS1 metaphase arrest is Mad1- and Mad2-dependent. Quantitation of the % large-budded cells with a single, DAPI-stained nucleus in the bud at the 3 hour time point (experiment repeated 3 times and >100 cells counted per sample). (E) Immunoblot of Myc-Mps1 levels in wild-type and mad mutants. Note that the Mps1 protein has reduced gel-mobility in the wild-type extract as these cells are arrested in mitosis where Mps1p is heavily phosphorylated. The myc tag was detected here using the 9B11 monoclonal. * indicates a non-specific band used as a loading control.
    Figure Legend Snippet: (A) P GAL7 -MPS1 was induced for 5 hours with 2% Galactose, or 2% glucose as the un-induced control. Scale bar is 10μm. (B) Quantitation of this mitotic arrest. Short spindles were scored every hour in at least 100 cells per condition at each time point. This experiment was repeated 3 times and displayed here as the mean +/- confidence levels. (C) Immunoblot of Mps1 levels at each hourly time point after Gal induction. Whole cell extracts were prepared by bead-beating, separated by SDS-PAGE and immunoblotted with the 9E10 anti-myc monoclonal antibody. * indicates a cross-reacting band, used here as a loading control. (D) P GAL7 -MPS1 metaphase arrest is Mad1- and Mad2-dependent. Quantitation of the % large-budded cells with a single, DAPI-stained nucleus in the bud at the 3 hour time point (experiment repeated 3 times and >100 cells counted per sample). (E) Immunoblot of Myc-Mps1 levels in wild-type and mad mutants. Note that the Mps1 protein has reduced gel-mobility in the wild-type extract as these cells are arrested in mitosis where Mps1p is heavily phosphorylated. The myc tag was detected here using the 9B11 monoclonal. * indicates a non-specific band used as a loading control.

    Techniques Used: Quantitation Assay, Western Blot, SDS Page, Staining

    (A) Mps1 in vitro kinase assays phosphorylate the Mad1 C-terminus. His-MBP is used here as a negative (specificity) control. Mps1 autophosphorylates and phosphorylates the C-terminus of Mad1. See for phosphopeptides identified in CMad1. (B) Quantitation of the phosphorylation of the Mad1 C-terminus. The radioactive Mad1 signal here was normalised to the Mad1 Coomassie band. (C) Alphafold model of the CnMad1-CTD compared to the crystal structure of hsMad1-CTD. Relevant threonine residues are highlighted on the model. (D) The mad1-T667A mutant, and the double (667,668) phospho-mutant, is benomyl sensitive. The strains indicated were grown at 30°C for 3 days. (E) Anti-GFP immunoblot of whole cell extracts. The GFP- mad1 phospho-mutant proteins are stable. See for a related immunoblot using the anti-Mad1 antibody. (F) The mad1-T667A mutant, and the double phospho-mutant, fails to nocodazole arrest. Cultures were grown to log phase and then challenged with nocodazole for 3 hours. This experiment was repeated 3 times. (G) The mad1-T667A mutant, and the double phospho-mutant, fails to arrest when Mps1 kinase is over-expressed. Log phase cultures were induced with 2% galactose for 5 hours. This experiment was repeated 3 times.
    Figure Legend Snippet: (A) Mps1 in vitro kinase assays phosphorylate the Mad1 C-terminus. His-MBP is used here as a negative (specificity) control. Mps1 autophosphorylates and phosphorylates the C-terminus of Mad1. See for phosphopeptides identified in CMad1. (B) Quantitation of the phosphorylation of the Mad1 C-terminus. The radioactive Mad1 signal here was normalised to the Mad1 Coomassie band. (C) Alphafold model of the CnMad1-CTD compared to the crystal structure of hsMad1-CTD. Relevant threonine residues are highlighted on the model. (D) The mad1-T667A mutant, and the double (667,668) phospho-mutant, is benomyl sensitive. The strains indicated were grown at 30°C for 3 days. (E) Anti-GFP immunoblot of whole cell extracts. The GFP- mad1 phospho-mutant proteins are stable. See for a related immunoblot using the anti-Mad1 antibody. (F) The mad1-T667A mutant, and the double phospho-mutant, fails to nocodazole arrest. Cultures were grown to log phase and then challenged with nocodazole for 3 hours. This experiment was repeated 3 times. (G) The mad1-T667A mutant, and the double phospho-mutant, fails to arrest when Mps1 kinase is over-expressed. Log phase cultures were induced with 2% galactose for 5 hours. This experiment was repeated 3 times.

    Techniques Used: In Vitro, Quantitation Assay, Mutagenesis, Western Blot

    For the full list of strains generated in this study.
    Figure Legend Snippet: For the full list of strains generated in this study.

    Techniques Used: Generated, Selection

    Primers used in this study.
    Figure Legend Snippet: Primers used in this study.

    Techniques Used: Sequencing, Clone Assay, Mutagenesis, Expressing

    Plasmids generated in this study.
    Figure Legend Snippet: Plasmids generated in this study.

    Techniques Used: Generated

    gfp monoclonal antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gfp monoclonal antibody
    Gfp Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp monoclonal antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    gfp monoclonal antibody - by Bioz Stars, 2024-07
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    antibodies against gfp  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc antibodies against gfp
    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected <t>GFP-tagged</t> <t>VAP</t> isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.
    Antibodies Against Gfp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "PITPβ promotes COPI vesicle fission through lipid transfer and membrane contact formation"

    Article Title: PITPβ promotes COPI vesicle fission through lipid transfer and membrane contact formation

    Journal: bioRxiv

    doi: 10.1101/2024.05.27.596058

    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected GFP-tagged VAP isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.
    Figure Legend Snippet: Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected GFP-tagged VAP isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.

    Techniques Used: Two Tailed Test, Transfection, Purification, Transport Assay, Quantitation Assay

    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, *** p<0.001. A , Co-precipitation experiment assessing the association of transfected HA-tagged PLD2 with transfected GFP-tagged PITPβ in cells, n=5. B , Co-precipitation experiment assessing the association of transfected HA-tagged LPP3 with transfected GFP-tagged PITPβ in cells, n=5. C , Dot blot analysis revealing that PITPβ binds directly to PC: PC, phosphatidylcholine; PA, phosphatidic acid; PS, phosphatidylserine; PE, phosphatidylethanolamine; Chol, cholesterol; SM, sphingomyelin; DAG, diacylglycerol; PI(4)P, phosphatidylinositol 4-phosphate, n= 4. D , Liposome binding study involving PITPβ incubated with liposome containing different lipids as indicated, n=5. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. E , Liposome binding study involving PITPβ incubated with Golgi-like liposomes, either having the full lipid composition or lacking a particular lipid as indicated, n=3. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. F , Dot blot analysis revealing that the catalytic mutant of PITPβ binds directly to PC. G , PITPβ can bind simultaneously to PA-containing liposomes, coatomer, and soluble VAP-A, n=3. H , Proximity ligation assay examining the effect of siRNA against PLD2 on the proximity between PITPβ and ζ−COP, n=4. I , Proximity ligation assay examining the effect of siRNA against LPP3 on the proximity between PITPβ and ζ−COP, n=4.
    Figure Legend Snippet: Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, *** p<0.001. A , Co-precipitation experiment assessing the association of transfected HA-tagged PLD2 with transfected GFP-tagged PITPβ in cells, n=5. B , Co-precipitation experiment assessing the association of transfected HA-tagged LPP3 with transfected GFP-tagged PITPβ in cells, n=5. C , Dot blot analysis revealing that PITPβ binds directly to PC: PC, phosphatidylcholine; PA, phosphatidic acid; PS, phosphatidylserine; PE, phosphatidylethanolamine; Chol, cholesterol; SM, sphingomyelin; DAG, diacylglycerol; PI(4)P, phosphatidylinositol 4-phosphate, n= 4. D , Liposome binding study involving PITPβ incubated with liposome containing different lipids as indicated, n=5. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. E , Liposome binding study involving PITPβ incubated with Golgi-like liposomes, either having the full lipid composition or lacking a particular lipid as indicated, n=3. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. F , Dot blot analysis revealing that the catalytic mutant of PITPβ binds directly to PC. G , PITPβ can bind simultaneously to PA-containing liposomes, coatomer, and soluble VAP-A, n=3. H , Proximity ligation assay examining the effect of siRNA against PLD2 on the proximity between PITPβ and ζ−COP, n=4. I , Proximity ligation assay examining the effect of siRNA against LPP3 on the proximity between PITPβ and ζ−COP, n=4.

    Techniques Used: Two Tailed Test, Transfection, Dot Blot, Binding Assay, Incubation, Membrane, Liposomes, Mutagenesis, Proximity Ligation Assay

    monoclonal anti gfp antibody b 2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc monoclonal anti gfp antibody b 2
    Monoclonal Anti Gfp Antibody B 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti gfp antibodies  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti gfp antibodies
    Anti Gfp Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc gfp antibody
    (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions <t>containing</t> <t>EphA2-GFP</t> were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.
    Gfp Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions <t>containing</t> <t>EphA2-GFP</t> were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.
    Antibodies Rabbit Anti Stim1 Mab D88e10 Cell Signaling 5668 Chicken Anti Gfp Abcam Ab13970, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions <t>containing</t> <t>EphA2-GFP</t> were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.
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    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected <t>GFP-tagged</t> <t>VAP</t> isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.
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    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected <t>GFP-tagged</t> <t>VAP</t> isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.
    Monoclonal Anti Gfp Antibody B 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected <t>GFP-tagged</t> <t>VAP</t> isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.
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    Image Search Results


    (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions containing EphA2-GFP were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.

    Journal: bioRxiv

    Article Title: Cholesterol inhibits assembly and activation of the EphA2 receptor

    doi: 10.1101/2024.06.10.598255

    Figure Lengend Snippet: (A) Schematic representation of sample preparation and workflow for SiMPull-POP. 1-Membrane fractions containing EphA2-GFP were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2-EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3-DIBMALPs devoid of EphA2-GFP are washed away before imaging. (B) Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody. Each blue spot represents a DIBMALP containing EphA2-GFP. (C) Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.

    Article Snippet: To immobilize protein samples, 0.02 mg/mL NeutrAvidin protein (Thermo Fisher Scientific) was first incubated in the chamber for 10 minutes followed by the addition of a biotinylated EphA2 or GFP antibody for 20 minutes (Cell Signaling and Rockland Immunochemical Inc., respectively).

    Techniques: Sample Prep, Membrane, Microscopy, Imaging

    (A) Schematic of DIBMALP containing an EphA2-GFP monomer. (B) Step distribution of control DIBMALPs (black) or those formed from cells treated with EA1 (pink), MβCD (blue) or both (magenta). (C) Oligomeric distribution calculated from data in panel B. (D) Schematic representing DDM micelles containing EphA2-GFP. (E) Step distribution of DDM-solubilized EphA2-GFP in the same conditions as in DIBMALPs. (F) Oligomeric distribution of DDM-solubilized EphA2-GFP photobleaching data. p -values are from two-way ANOVA followed by Tukey multiple comparison test.*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

    Journal: bioRxiv

    Article Title: Cholesterol inhibits assembly and activation of the EphA2 receptor

    doi: 10.1101/2024.06.10.598255

    Figure Lengend Snippet: (A) Schematic of DIBMALP containing an EphA2-GFP monomer. (B) Step distribution of control DIBMALPs (black) or those formed from cells treated with EA1 (pink), MβCD (blue) or both (magenta). (C) Oligomeric distribution calculated from data in panel B. (D) Schematic representing DDM micelles containing EphA2-GFP. (E) Step distribution of DDM-solubilized EphA2-GFP in the same conditions as in DIBMALPs. (F) Oligomeric distribution of DDM-solubilized EphA2-GFP photobleaching data. p -values are from two-way ANOVA followed by Tukey multiple comparison test.*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

    Article Snippet: To immobilize protein samples, 0.02 mg/mL NeutrAvidin protein (Thermo Fisher Scientific) was first incubated in the chamber for 10 minutes followed by the addition of a biotinylated EphA2 or GFP antibody for 20 minutes (Cell Signaling and Rockland Immunochemical Inc., respectively).

    Techniques: Comparison

    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected GFP-tagged VAP isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.

    Journal: bioRxiv

    Article Title: PITPβ promotes COPI vesicle fission through lipid transfer and membrane contact formation

    doi: 10.1101/2024.05.27.596058

    Figure Lengend Snippet: Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, ns p>0.05. A , Co-precipitation experiment assessing the association of transfected GFP-tagged VAP isoforms with endogenous PITPβ in cells, n=6. B , Pulldown experiment using purified components to assess the effect of mutating dilysine motifs in PITPβ on its direct interaction with coatomer, n=5. C , COPI transport assay assessing the effect of mutating the FFAT motif in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm. D , Pulldown experiment using purified components to confirm a specific dilysine motif in full-length PITPβ is critical for its direct interaction with coatomer, n=5. E , COPI transport assay assessing the effect of mutating dilysine sequences in PITPβ. Quantitation is shown on right, n=5. Representative confocal images are shown on left, with VSVG-KDELR colored in red and giantin colored in green, bar=10 µm.

    Article Snippet: The following antibodies were obtained: monoclonal antibody against PITPβ (kind gift from Dr. Shamshad Cockcroft, University College London); anti-HA epitope (3724S), and 6xhis epitope antibodies (9991S) from Cell Signaling; antibodies against GFP (sc-9996), VAP-A (sc-293278), calnexin (sc-46669), and β-actin (sc-47778) from Santa Cruz Biotechnology; antibodies against anti-GM130 (610822) from BD Transduction Laboratories: mouse anti-LPP3 (ab52581) and rabbit anti-giantin (ab80864) antibodies from Abcam; rabbit anti-TGN46 (PA1-1069) and anti-ST6GAL1 (PA5-106647) antibodies from Invitrogen.

    Techniques: Two Tailed Test, Transfection, Purification, Transport Assay, Quantitation Assay

    Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, *** p<0.001. A , Co-precipitation experiment assessing the association of transfected HA-tagged PLD2 with transfected GFP-tagged PITPβ in cells, n=5. B , Co-precipitation experiment assessing the association of transfected HA-tagged LPP3 with transfected GFP-tagged PITPβ in cells, n=5. C , Dot blot analysis revealing that PITPβ binds directly to PC: PC, phosphatidylcholine; PA, phosphatidic acid; PS, phosphatidylserine; PE, phosphatidylethanolamine; Chol, cholesterol; SM, sphingomyelin; DAG, diacylglycerol; PI(4)P, phosphatidylinositol 4-phosphate, n= 4. D , Liposome binding study involving PITPβ incubated with liposome containing different lipids as indicated, n=5. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. E , Liposome binding study involving PITPβ incubated with Golgi-like liposomes, either having the full lipid composition or lacking a particular lipid as indicated, n=3. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. F , Dot blot analysis revealing that the catalytic mutant of PITPβ binds directly to PC. G , PITPβ can bind simultaneously to PA-containing liposomes, coatomer, and soluble VAP-A, n=3. H , Proximity ligation assay examining the effect of siRNA against PLD2 on the proximity between PITPβ and ζ−COP, n=4. I , Proximity ligation assay examining the effect of siRNA against LPP3 on the proximity between PITPβ and ζ−COP, n=4.

    Journal: bioRxiv

    Article Title: PITPβ promotes COPI vesicle fission through lipid transfer and membrane contact formation

    doi: 10.1101/2024.05.27.596058

    Figure Lengend Snippet: Quantitative data are shown as mean ± s.e.m, with the number of independent experiments indicated. Statistics was performed using the two-tailed student t -test: **** p<0.0001, *** p<0.001. A , Co-precipitation experiment assessing the association of transfected HA-tagged PLD2 with transfected GFP-tagged PITPβ in cells, n=5. B , Co-precipitation experiment assessing the association of transfected HA-tagged LPP3 with transfected GFP-tagged PITPβ in cells, n=5. C , Dot blot analysis revealing that PITPβ binds directly to PC: PC, phosphatidylcholine; PA, phosphatidic acid; PS, phosphatidylserine; PE, phosphatidylethanolamine; Chol, cholesterol; SM, sphingomyelin; DAG, diacylglycerol; PI(4)P, phosphatidylinositol 4-phosphate, n= 4. D , Liposome binding study involving PITPβ incubated with liposome containing different lipids as indicated, n=5. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. E , Liposome binding study involving PITPβ incubated with Golgi-like liposomes, either having the full lipid composition or lacking a particular lipid as indicated, n=3. Pellet (P) fraction contains membrane-bound PITPβ, while supernatant (S) fraction contains soluble PITPβ. F , Dot blot analysis revealing that the catalytic mutant of PITPβ binds directly to PC. G , PITPβ can bind simultaneously to PA-containing liposomes, coatomer, and soluble VAP-A, n=3. H , Proximity ligation assay examining the effect of siRNA against PLD2 on the proximity between PITPβ and ζ−COP, n=4. I , Proximity ligation assay examining the effect of siRNA against LPP3 on the proximity between PITPβ and ζ−COP, n=4.

    Article Snippet: The following antibodies were obtained: monoclonal antibody against PITPβ (kind gift from Dr. Shamshad Cockcroft, University College London); anti-HA epitope (3724S), and 6xhis epitope antibodies (9991S) from Cell Signaling; antibodies against GFP (sc-9996), VAP-A (sc-293278), calnexin (sc-46669), and β-actin (sc-47778) from Santa Cruz Biotechnology; antibodies against anti-GM130 (610822) from BD Transduction Laboratories: mouse anti-LPP3 (ab52581) and rabbit anti-giantin (ab80864) antibodies from Abcam; rabbit anti-TGN46 (PA1-1069) and anti-ST6GAL1 (PA5-106647) antibodies from Invitrogen.

    Techniques: Two Tailed Test, Transfection, Dot Blot, Binding Assay, Incubation, Membrane, Liposomes, Mutagenesis, Proximity Ligation Assay