monoclonal anti flag m2 antibody Millipore Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore anti flag mab
    Anti Flag Mab, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 539 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag mab/product/Millipore
    Average 99 stars, based on 539 article reviews
    Price from $9.99 to $1999.99
    anti flag mab - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

    91
    Millipore m2 anti flag mab
    M2 Anti Flag Mab, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 459 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m2 anti flag mab/product/Millipore
    Average 91 stars, based on 459 article reviews
    Price from $9.99 to $1999.99
    m2 anti flag mab - by Bioz Stars, 2020-01
    91/100 stars
      Buy from Supplier

    76
    Millipore m2 flag mabs
    M2 Flag Mabs, supplied by Millipore, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m2 flag mabs/product/Millipore
    Average 76 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    m2 flag mabs - by Bioz Stars, 2020-01
    76/100 stars
      Buy from Supplier

    79
    Millipore anti flag m2 monoclonal ab mab
    Anti Flag M2 Monoclonal Ab Mab, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag m2 monoclonal ab mab/product/Millipore
    Average 79 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    anti flag m2 monoclonal ab mab - by Bioz Stars, 2020-01
    79/100 stars
      Buy from Supplier

    94
    Millipore m2 anti flag mab agarose beads
    M2 Anti Flag Mab Agarose Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m2 anti flag mab agarose beads/product/Millipore
    Average 94 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    m2 anti flag mab agarose beads - by Bioz Stars, 2020-01
    94/100 stars
      Buy from Supplier

    77
    Millipore anti fluoresceinated antigen flag m2 mab
    Anti Fluoresceinated Antigen Flag M2 Mab, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti fluoresceinated antigen flag m2 mab/product/Millipore
    Average 77 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    anti fluoresceinated antigen flag m2 mab - by Bioz Stars, 2020-01
    77/100 stars
      Buy from Supplier

    78
    Millipore anti flag bio m2 mab
    Anti Flag Bio M2 Mab, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag bio m2 mab/product/Millipore
    Average 78 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    anti flag bio m2 mab - by Bioz Stars, 2020-01
    78/100 stars
      Buy from Supplier

    79
    Millipore anti flag m2 horseradish peroxidase hrp mab mab 1 1 000
    Anti Flag M2 Horseradish Peroxidase Hrp Mab Mab 1 1 000, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag m2 horseradish peroxidase hrp mab mab 1 1 000/product/Millipore
    Average 79 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    anti flag m2 horseradish peroxidase hrp mab mab 1 1 000 - by Bioz Stars, 2020-01
    79/100 stars
      Buy from Supplier

    79
    Millipore anti flag mab m2 fitc
    Domain architecture of the scFv:G28 fusion proteins used in this study and initial analysis of scFv:G28-TRAIL. ( a ) Schemes of scFv:G28-TRAIL, scFv:G28-2xFlag-GpL, scFv:G28-Fc-GpL and scFv:G28-TNC-GpL. scFv:G28, scFv derived from the human CD40-specific mAb G28-5; Flag epitope; TNC, trimerization domain, aa 110-139 of chicken tenascin-C; TRAIL, aa 95-281 of human TRAIL; Fc, CH3 domain of hIgG1; GpL, luciferase of Gaussia princeps . scFv:G28-Fc-GpL and scFv:G28-2xFlag-GpL were used as competitors for scFv:G28-TRAIL. scFv:G28-TNC-GpL were used to analyze the effect of trimerization of scFv:G28 on CD40 activity. ( b ) Anti-Flag affinity chromatography purified scFv:G28-TRAIL (100 ng) was subjected to SDS-PAGE and visualized by silver staining. ( c ) Jurkat cells (60 × 10 3 per well; 96-well plate) were challenged in the absence and presence of the anti-Flag mAb M2 (1 μ g/ml) with the indicated concentrations of scFv:G28-TRAIL or its conventional counterpart Flag-TNC-TRAIL. Next day, cellular viability was determined by using the MTT assay ( d ) HeLa and HT1080 cells along with the corresponding CD40 transfectants were incubated with scFv:G28-TRAIL (500 ng/ml) and after three washes, bound molecules were detected by FACS using anti-Flag mAb <t>M2-FITC</t>
    Anti Flag Mab M2 Fitc, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag mab m2 fitc/product/Millipore
    Average 79 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    anti flag mab m2 fitc - by Bioz Stars, 2020-01
    79/100 stars
      Buy from Supplier

    86
    Merck KGaA anti flag m2 monoclonal
    Domain architecture of the scFv:G28 fusion proteins used in this study and initial analysis of scFv:G28-TRAIL. ( a ) Schemes of scFv:G28-TRAIL, scFv:G28-2xFlag-GpL, scFv:G28-Fc-GpL and scFv:G28-TNC-GpL. scFv:G28, scFv derived from the human CD40-specific mAb G28-5; Flag epitope; TNC, trimerization domain, aa 110-139 of chicken tenascin-C; TRAIL, aa 95-281 of human TRAIL; Fc, CH3 domain of hIgG1; GpL, luciferase of Gaussia princeps . scFv:G28-Fc-GpL and scFv:G28-2xFlag-GpL were used as competitors for scFv:G28-TRAIL. scFv:G28-TNC-GpL were used to analyze the effect of trimerization of scFv:G28 on CD40 activity. ( b ) Anti-Flag affinity chromatography purified scFv:G28-TRAIL (100 ng) was subjected to SDS-PAGE and visualized by silver staining. ( c ) Jurkat cells (60 × 10 3 per well; 96-well plate) were challenged in the absence and presence of the anti-Flag mAb M2 (1 μ g/ml) with the indicated concentrations of scFv:G28-TRAIL or its conventional counterpart Flag-TNC-TRAIL. Next day, cellular viability was determined by using the MTT assay ( d ) HeLa and HT1080 cells along with the corresponding CD40 transfectants were incubated with scFv:G28-TRAIL (500 ng/ml) and after three washes, bound molecules were detected by FACS using anti-Flag mAb <t>M2-FITC</t>
    Anti Flag M2 Monoclonal, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag m2 monoclonal/product/Merck KGaA
    Average 86 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    anti flag m2 monoclonal - by Bioz Stars, 2020-01
    86/100 stars
      Buy from Supplier

    99
    Millipore anti flag monoclonal antibody
    Domain architecture of the scFv:G28 fusion proteins used in this study and initial analysis of scFv:G28-TRAIL. ( a ) Schemes of scFv:G28-TRAIL, scFv:G28-2xFlag-GpL, scFv:G28-Fc-GpL and scFv:G28-TNC-GpL. scFv:G28, scFv derived from the human CD40-specific mAb G28-5; Flag epitope; TNC, trimerization domain, aa 110-139 of chicken tenascin-C; TRAIL, aa 95-281 of human TRAIL; Fc, CH3 domain of hIgG1; GpL, luciferase of Gaussia princeps . scFv:G28-Fc-GpL and scFv:G28-2xFlag-GpL were used as competitors for scFv:G28-TRAIL. scFv:G28-TNC-GpL were used to analyze the effect of trimerization of scFv:G28 on CD40 activity. ( b ) Anti-Flag affinity chromatography purified scFv:G28-TRAIL (100 ng) was subjected to SDS-PAGE and visualized by silver staining. ( c ) Jurkat cells (60 × 10 3 per well; 96-well plate) were challenged in the absence and presence of the anti-Flag mAb M2 (1 μ g/ml) with the indicated concentrations of scFv:G28-TRAIL or its conventional counterpart Flag-TNC-TRAIL. Next day, cellular viability was determined by using the MTT assay ( d ) HeLa and HT1080 cells along with the corresponding CD40 transfectants were incubated with scFv:G28-TRAIL (500 ng/ml) and after three washes, bound molecules were detected by FACS using anti-Flag mAb <t>M2-FITC</t>
    Anti Flag Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1491 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag monoclonal antibody/product/Millipore
    Average 99 stars, based on 1491 article reviews
    Price from $9.99 to $1999.99
    anti flag monoclonal antibody - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

    99
    Millipore monoclonal anti flag m2 antibody
    Expression of Ta3FL, but not Ta3 mutants, rescues ciliogenesis in Ta3 mutant cells. ( A–G” ) Ta3 mutant MEFs were transduced with the retrovirus carrying the constructs indicated above the panels. Following serum starvation, the cells were stained with <t>FLAG,</t> Cep120, and Arl13b <t>antibodies</t> and counterstained with DAPI for nuclei. FLAG labels Ta3 and mutants. Cep120 marks centrioles, and Arl13b is a cilia marker. Insets are enlargement of the framed areas with dash lines. Note that although both FH-Ta3FL and FH-Ta3-1–638 localize at centrioles, only FH-Ta3FL expression rescues cilia formation. ( H ) A graph showing that the percent cells that are FLAG-staining positive at centrioles form cilia. Two-tailed Student t -test p-value = 0.0001 (n=72). ( I ) Immunoblots showing the expression of Ta3 and mutant proteins.
    Monoclonal Anti Flag M2 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 29200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti flag m2 antibody/product/Millipore
    Average 99 stars, based on 29200 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti flag m2 antibody - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

    99
    Millipore mouse monoclonal anti flag
    Expression of Ta3FL, but not Ta3 mutants, rescues ciliogenesis in Ta3 mutant cells. ( A–G” ) Ta3 mutant MEFs were transduced with the retrovirus carrying the constructs indicated above the panels. Following serum starvation, the cells were stained with <t>FLAG,</t> Cep120, and Arl13b <t>antibodies</t> and counterstained with DAPI for nuclei. FLAG labels Ta3 and mutants. Cep120 marks centrioles, and Arl13b is a cilia marker. Insets are enlargement of the framed areas with dash lines. Note that although both FH-Ta3FL and FH-Ta3-1–638 localize at centrioles, only FH-Ta3FL expression rescues cilia formation. ( H ) A graph showing that the percent cells that are FLAG-staining positive at centrioles form cilia. Two-tailed Student t -test p-value = 0.0001 (n=72). ( I ) Immunoblots showing the expression of Ta3 and mutant proteins.
    Mouse Monoclonal Anti Flag, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1028 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti flag/product/Millipore
    Average 99 stars, based on 1028 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti flag - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

    87
    Millipore flag monoclonal antibody mab ip
    STAU1 directly interacts with human PCP2 mRNA. a Non-RNase A treated HEK-293 whole-cell extracts expressing <t>Flag</t> or Flag-STAU1 were subjected to immunoprecipitation with Flag <t>mAb</t> beads. Flag-STAU1 pulled down ATXN2 protein, and PCP2 and CALB1 mRNAs on western blot and RT-PCR analysis. b – e Northwestern blotting showing that STAU1 binds directly to human PCP2 RNA in a manner requiring the 3′UTR. Construction of DIG-labeled human PCP2 RNA probes with both UTRs ( b ), 3′ UTR only ( c ), and 5′UTR only ( d ). e Bacterial expressed His-STAU1, His-GFP, or control bacterial lysate (vector alone) were run on SDS–PAGE and stained with Coomassie brilliant blue (CBB) or western blotted with anti-His <t>antibody</t> (blot 1). Protein blots were hybridized with DIG-labeled PCP2 RNA probes followed by anti-DIG antibody staining. STAU1 directly interacts with PCP2 RNAs: ( 5′ + 3′)UTRs (blot 2) or 3′UTR (blot 3), but not with PCP2(5′UTR) RNA (blot 4). His-GFP shows no interactions. The blot represents one of three independent experiments. f STAU1- PCP2 RNA granules in SCA2 cells. Normal and SCA2 FBs were immunostained with STAU1 antibodies and STAU1 granules (green) are visualized in SCA2 FBs but not in normal FBs. PCP2 RNA granules (red) in SCA2 FBs were detected by FISH using a PCP2 -Cy3 probe. Merged images demonstrate positive STAU1- PCP2 RNA granules (white arrows) for SCA2 FBs. Scale bar, 30 µM
    Flag Monoclonal Antibody Mab Ip, supplied by Millipore, used in various techniques. Bioz Stars score: 87/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flag monoclonal antibody mab ip/product/Millipore
    Average 87 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    flag monoclonal antibody mab ip - by Bioz Stars, 2020-01
    87/100 stars
      Buy from Supplier

    83
    Millipore flag specific mab m2 coupled agarose beads
    STAU1 directly interacts with human PCP2 mRNA. a Non-RNase A treated HEK-293 whole-cell extracts expressing <t>Flag</t> or Flag-STAU1 were subjected to immunoprecipitation with Flag <t>mAb</t> beads. Flag-STAU1 pulled down ATXN2 protein, and PCP2 and CALB1 mRNAs on western blot and RT-PCR analysis. b – e Northwestern blotting showing that STAU1 binds directly to human PCP2 RNA in a manner requiring the 3′UTR. Construction of DIG-labeled human PCP2 RNA probes with both UTRs ( b ), 3′ UTR only ( c ), and 5′UTR only ( d ). e Bacterial expressed His-STAU1, His-GFP, or control bacterial lysate (vector alone) were run on SDS–PAGE and stained with Coomassie brilliant blue (CBB) or western blotted with anti-His <t>antibody</t> (blot 1). Protein blots were hybridized with DIG-labeled PCP2 RNA probes followed by anti-DIG antibody staining. STAU1 directly interacts with PCP2 RNAs: ( 5′ + 3′)UTRs (blot 2) or 3′UTR (blot 3), but not with PCP2(5′UTR) RNA (blot 4). His-GFP shows no interactions. The blot represents one of three independent experiments. f STAU1- PCP2 RNA granules in SCA2 cells. Normal and SCA2 FBs were immunostained with STAU1 antibodies and STAU1 granules (green) are visualized in SCA2 FBs but not in normal FBs. PCP2 RNA granules (red) in SCA2 FBs were detected by FISH using a PCP2 -Cy3 probe. Merged images demonstrate positive STAU1- PCP2 RNA granules (white arrows) for SCA2 FBs. Scale bar, 30 µM
    Flag Specific Mab M2 Coupled Agarose Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 83/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flag specific mab m2 coupled agarose beads/product/Millipore
    Average 83 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    flag specific mab m2 coupled agarose beads - by Bioz Stars, 2020-01
    83/100 stars
      Buy from Supplier

    98
    Millipore monoclonal antibody
    STAU1 directly interacts with human PCP2 mRNA. a Non-RNase A treated HEK-293 whole-cell extracts expressing <t>Flag</t> or Flag-STAU1 were subjected to immunoprecipitation with Flag <t>mAb</t> beads. Flag-STAU1 pulled down ATXN2 protein, and PCP2 and CALB1 mRNAs on western blot and RT-PCR analysis. b – e Northwestern blotting showing that STAU1 binds directly to human PCP2 RNA in a manner requiring the 3′UTR. Construction of DIG-labeled human PCP2 RNA probes with both UTRs ( b ), 3′ UTR only ( c ), and 5′UTR only ( d ). e Bacterial expressed His-STAU1, His-GFP, or control bacterial lysate (vector alone) were run on SDS–PAGE and stained with Coomassie brilliant blue (CBB) or western blotted with anti-His <t>antibody</t> (blot 1). Protein blots were hybridized with DIG-labeled PCP2 RNA probes followed by anti-DIG antibody staining. STAU1 directly interacts with PCP2 RNAs: ( 5′ + 3′)UTRs (blot 2) or 3′UTR (blot 3), but not with PCP2(5′UTR) RNA (blot 4). His-GFP shows no interactions. The blot represents one of three independent experiments. f STAU1- PCP2 RNA granules in SCA2 cells. Normal and SCA2 FBs were immunostained with STAU1 antibodies and STAU1 granules (green) are visualized in SCA2 FBs but not in normal FBs. PCP2 RNA granules (red) in SCA2 FBs were detected by FISH using a PCP2 -Cy3 probe. Merged images demonstrate positive STAU1- PCP2 RNA granules (white arrows) for SCA2 FBs. Scale bar, 30 µM
    Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1546 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal antibody/product/Millipore
    Average 98 stars, based on 1546 article reviews
    Price from $9.99 to $1999.99
    monoclonal antibody - by Bioz Stars, 2020-01
    98/100 stars
      Buy from Supplier

    99
    Millipore monoclonal anti flag biom2 antibody
    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with <t>anti-FLAG,</t> anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups
    Monoclonal Anti Flag Biom2 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 247 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti flag biom2 antibody/product/Millipore
    Average 99 stars, based on 247 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti flag biom2 antibody - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

    78
    Millipore monoclonal mouse anti flag epitope
    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with <t>anti-FLAG,</t> anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups
    Monoclonal Mouse Anti Flag Epitope, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal mouse anti flag epitope/product/Millipore
    Average 78 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    monoclonal mouse anti flag epitope - by Bioz Stars, 2020-01
    78/100 stars
      Buy from Supplier

    99
    Millipore mouse monoclonal anti flag m2
    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with <t>anti-FLAG,</t> anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups
    Mouse Monoclonal Anti Flag M2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1050 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti flag m2/product/Millipore
    Average 99 stars, based on 1050 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti flag m2 - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

    78
    Millipore mouse monoclonal m2 anti 3
    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with <t>anti-FLAG,</t> anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups
    Mouse Monoclonal M2 Anti 3, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal m2 anti 3/product/Millipore
    Average 78 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal m2 anti 3 - by Bioz Stars, 2020-01
    78/100 stars
      Buy from Supplier

    70
    Millipore monoclonal anti flag m2 agarose
    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with <t>anti-FLAG,</t> anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups
    Monoclonal Anti Flag M2 Agarose, supplied by Millipore, used in various techniques. Bioz Stars score: 70/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti flag m2 agarose/product/Millipore
    Average 70 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti flag m2 agarose - by Bioz Stars, 2020-01
    70/100 stars
      Buy from Supplier

    88
    Millipore anti flag m2 monoclonal igg
    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with <t>anti-FLAG,</t> anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups
    Anti Flag M2 Monoclonal Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag m2 monoclonal igg/product/Millipore
    Average 88 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    anti flag m2 monoclonal igg - by Bioz Stars, 2020-01
    88/100 stars
      Buy from Supplier

    94
    Millipore monoclonal anti flag m2 cy3 antibody
    Subcellular localization of <t>TIG3-FLAG</t> fusion proteins. A Schematic of the fusion proteins in which the segments of the N-terminal hydrophilic region are fused to FLAG epitope. The TIG3 domains are described in the legend to Fig. 1A . B SCC-13 cells were electroporated with 3 µg of each TIG3-FLAG fusion protein and after 24 h the cells were fixed and co-stained with the centrosome marker, pericentrin (green), and <t>Cy3-conjugated</t> <t>anti-FLAG</t> (red). Each of these fusion proteins localize at the centrosome (yellow, arrows). Bars = 10 µm. C TIG3(102–125)-FLAG accumulates at the centrosome in A431 cells and normal foreskin keratinocytes (KERn). Cells were electroporated with plasmid encoding TIG3(102–125)-FLAG and then stained with antibodies as indicated above. Bars = 10 µM.
    Monoclonal Anti Flag M2 Cy3 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti flag m2 cy3 antibody/product/Millipore
    Average 94 stars, based on 73 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti flag m2 cy3 antibody - by Bioz Stars, 2020-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    Domain architecture of the scFv:G28 fusion proteins used in this study and initial analysis of scFv:G28-TRAIL. ( a ) Schemes of scFv:G28-TRAIL, scFv:G28-2xFlag-GpL, scFv:G28-Fc-GpL and scFv:G28-TNC-GpL. scFv:G28, scFv derived from the human CD40-specific mAb G28-5; Flag epitope; TNC, trimerization domain, aa 110-139 of chicken tenascin-C; TRAIL, aa 95-281 of human TRAIL; Fc, CH3 domain of hIgG1; GpL, luciferase of Gaussia princeps . scFv:G28-Fc-GpL and scFv:G28-2xFlag-GpL were used as competitors for scFv:G28-TRAIL. scFv:G28-TNC-GpL were used to analyze the effect of trimerization of scFv:G28 on CD40 activity. ( b ) Anti-Flag affinity chromatography purified scFv:G28-TRAIL (100 ng) was subjected to SDS-PAGE and visualized by silver staining. ( c ) Jurkat cells (60 × 10 3 per well; 96-well plate) were challenged in the absence and presence of the anti-Flag mAb M2 (1 μ g/ml) with the indicated concentrations of scFv:G28-TRAIL or its conventional counterpart Flag-TNC-TRAIL. Next day, cellular viability was determined by using the MTT assay ( d ) HeLa and HT1080 cells along with the corresponding CD40 transfectants were incubated with scFv:G28-TRAIL (500 ng/ml) and after three washes, bound molecules were detected by FACS using anti-Flag mAb M2-FITC

    Journal: Cell Death & Disease

    Article Title: CD40-directed scFv-TRAIL fusion proteins induce CD40-restricted tumor cell death and activate dendritic cells

    doi: 10.1038/cddis.2013.402

    Figure Lengend Snippet: Domain architecture of the scFv:G28 fusion proteins used in this study and initial analysis of scFv:G28-TRAIL. ( a ) Schemes of scFv:G28-TRAIL, scFv:G28-2xFlag-GpL, scFv:G28-Fc-GpL and scFv:G28-TNC-GpL. scFv:G28, scFv derived from the human CD40-specific mAb G28-5; Flag epitope; TNC, trimerization domain, aa 110-139 of chicken tenascin-C; TRAIL, aa 95-281 of human TRAIL; Fc, CH3 domain of hIgG1; GpL, luciferase of Gaussia princeps . scFv:G28-Fc-GpL and scFv:G28-2xFlag-GpL were used as competitors for scFv:G28-TRAIL. scFv:G28-TNC-GpL were used to analyze the effect of trimerization of scFv:G28 on CD40 activity. ( b ) Anti-Flag affinity chromatography purified scFv:G28-TRAIL (100 ng) was subjected to SDS-PAGE and visualized by silver staining. ( c ) Jurkat cells (60 × 10 3 per well; 96-well plate) were challenged in the absence and presence of the anti-Flag mAb M2 (1 μ g/ml) with the indicated concentrations of scFv:G28-TRAIL or its conventional counterpart Flag-TNC-TRAIL. Next day, cellular viability was determined by using the MTT assay ( d ) HeLa and HT1080 cells along with the corresponding CD40 transfectants were incubated with scFv:G28-TRAIL (500 ng/ml) and after three washes, bound molecules were detected by FACS using anti-Flag mAb M2-FITC

    Article Snippet: Cells were washed again three times with PBS and incubated on ice for 30 min with anti-Flag mAb M2-FITC (Sigma-Aldrich).

    Techniques: Derivative Assay, FLAG-tag, Luciferase, Activity Assay, Affinity Chromatography, Purification, SDS Page, Silver Staining, MTT Assay, Incubation, FACS

    Expression of Ta3FL, but not Ta3 mutants, rescues ciliogenesis in Ta3 mutant cells. ( A–G” ) Ta3 mutant MEFs were transduced with the retrovirus carrying the constructs indicated above the panels. Following serum starvation, the cells were stained with FLAG, Cep120, and Arl13b antibodies and counterstained with DAPI for nuclei. FLAG labels Ta3 and mutants. Cep120 marks centrioles, and Arl13b is a cilia marker. Insets are enlargement of the framed areas with dash lines. Note that although both FH-Ta3FL and FH-Ta3-1–638 localize at centrioles, only FH-Ta3FL expression rescues cilia formation. ( H ) A graph showing that the percent cells that are FLAG-staining positive at centrioles form cilia. Two-tailed Student t -test p-value = 0.0001 (n=72). ( I ) Immunoblots showing the expression of Ta3 and mutant proteins.

    Journal: Developmental biology

    Article Title: PKA-mediated Gli2 and Gli3 phosphorylation is inhibited by Hedgehog signaling in cilia and reduced in Talpid3 mutant

    doi: 10.1016/j.ydbio.2017.06.035

    Figure Lengend Snippet: Expression of Ta3FL, but not Ta3 mutants, rescues ciliogenesis in Ta3 mutant cells. ( A–G” ) Ta3 mutant MEFs were transduced with the retrovirus carrying the constructs indicated above the panels. Following serum starvation, the cells were stained with FLAG, Cep120, and Arl13b antibodies and counterstained with DAPI for nuclei. FLAG labels Ta3 and mutants. Cep120 marks centrioles, and Arl13b is a cilia marker. Insets are enlargement of the framed areas with dash lines. Note that although both FH-Ta3FL and FH-Ta3-1–638 localize at centrioles, only FH-Ta3FL expression rescues cilia formation. ( H ) A graph showing that the percent cells that are FLAG-staining positive at centrioles form cilia. Two-tailed Student t -test p-value = 0.0001 (n=72). ( I ) Immunoblots showing the expression of Ta3 and mutant proteins.

    Article Snippet: Other antibodies include: Gli2 N-terminal, Gli3 N-terminal, Ta3, Cep120 (all 1:1000) ( ; ; ), acetylated tubulin (1:4000), FLAG (T6793, F1804, Sigma), PKARIIβ (610625, BD Biosciences), and Myc (sc-788, Santa Cruz Biotechnology).

    Techniques: Expressing, Mutagenesis, Transduction, Construct, Staining, Marker, Two Tailed Test, Western Blot

    STAU1 directly interacts with human PCP2 mRNA. a Non-RNase A treated HEK-293 whole-cell extracts expressing Flag or Flag-STAU1 were subjected to immunoprecipitation with Flag mAb beads. Flag-STAU1 pulled down ATXN2 protein, and PCP2 and CALB1 mRNAs on western blot and RT-PCR analysis. b – e Northwestern blotting showing that STAU1 binds directly to human PCP2 RNA in a manner requiring the 3′UTR. Construction of DIG-labeled human PCP2 RNA probes with both UTRs ( b ), 3′ UTR only ( c ), and 5′UTR only ( d ). e Bacterial expressed His-STAU1, His-GFP, or control bacterial lysate (vector alone) were run on SDS–PAGE and stained with Coomassie brilliant blue (CBB) or western blotted with anti-His antibody (blot 1). Protein blots were hybridized with DIG-labeled PCP2 RNA probes followed by anti-DIG antibody staining. STAU1 directly interacts with PCP2 RNAs: ( 5′ + 3′)UTRs (blot 2) or 3′UTR (blot 3), but not with PCP2(5′UTR) RNA (blot 4). His-GFP shows no interactions. The blot represents one of three independent experiments. f STAU1- PCP2 RNA granules in SCA2 cells. Normal and SCA2 FBs were immunostained with STAU1 antibodies and STAU1 granules (green) are visualized in SCA2 FBs but not in normal FBs. PCP2 RNA granules (red) in SCA2 FBs were detected by FISH using a PCP2 -Cy3 probe. Merged images demonstrate positive STAU1- PCP2 RNA granules (white arrows) for SCA2 FBs. Scale bar, 30 µM

    Journal: Nature Communications

    Article Title: Staufen1 links RNA stress granules and autophagy in a model of neurodegeneration

    doi: 10.1038/s41467-018-06041-3

    Figure Lengend Snippet: STAU1 directly interacts with human PCP2 mRNA. a Non-RNase A treated HEK-293 whole-cell extracts expressing Flag or Flag-STAU1 were subjected to immunoprecipitation with Flag mAb beads. Flag-STAU1 pulled down ATXN2 protein, and PCP2 and CALB1 mRNAs on western blot and RT-PCR analysis. b – e Northwestern blotting showing that STAU1 binds directly to human PCP2 RNA in a manner requiring the 3′UTR. Construction of DIG-labeled human PCP2 RNA probes with both UTRs ( b ), 3′ UTR only ( c ), and 5′UTR only ( d ). e Bacterial expressed His-STAU1, His-GFP, or control bacterial lysate (vector alone) were run on SDS–PAGE and stained with Coomassie brilliant blue (CBB) or western blotted with anti-His antibody (blot 1). Protein blots were hybridized with DIG-labeled PCP2 RNA probes followed by anti-DIG antibody staining. STAU1 directly interacts with PCP2 RNAs: ( 5′ + 3′)UTRs (blot 2) or 3′UTR (blot 3), but not with PCP2(5′UTR) RNA (blot 4). His-GFP shows no interactions. The blot represents one of three independent experiments. f STAU1- PCP2 RNA granules in SCA2 cells. Normal and SCA2 FBs were immunostained with STAU1 antibodies and STAU1 granules (green) are visualized in SCA2 FBs but not in normal FBs. PCP2 RNA granules (red) in SCA2 FBs were detected by FISH using a PCP2 -Cy3 probe. Merged images demonstrate positive STAU1- PCP2 RNA granules (white arrows) for SCA2 FBs. Scale bar, 30 µM

    Article Snippet: Ninety percent of cell extracts were subjected to Flag monoclonal antibody (mAb) IP (Anti-Flag M2 Affinity Gel, Sigma-Aldrich, A2220-1ML) to immunoprecipitate ATXN2 or STAU1 interacting protein–RNA complexes.

    Techniques: Expressing, Immunoprecipitation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Labeling, Plasmid Preparation, SDS Page, Staining, Northern blot, Fluorescence In Situ Hybridization

    PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with anti-FLAG, anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups

    Journal: Nature Communications

    Article Title: Functionally distinct and selectively phosphorylated GPCR subpopulations co-exist in a single cell

    doi: 10.1038/s41467-018-03459-7

    Figure Lengend Snippet: PKA- and GRK-pβ 2 ARs are spatially segregated on the plasma membrane. a , b SIM imaging shows total β 2 ARs, and PKA- and GRK-phosphorylated β 2 ARs, which were stained with anti-FLAG, anti-S261/262 (PKA-pβ 2 AR), and anti-S355/356 (GRK-pβ 2 AR) specific antibodies, respectively, in HEK293 cells expressing FLAG-tagged β 2 AR before stimulation ( a ) or after 30 s of stimulation with 1 μM ISO ( b ). Scale bar, 2 μm. Representative of n = 5, 4, 19, 11, and 13 cells, respectively, three independent experiments. c The overlap between two different staining from images in b was evaluated by Pearson’s correlation coefficient ( n = 19, 11, and 13 cells, respectively). d Quantification of two-color overlap was evaluated in HEK293 cells after 1 min of stimulation with 1 μM ISO using the same method in b ( n = 13, 12, and 11 cells, respectively, three independent experiments). e FLAG-β 2 ARs expressed in HEK293 cells were stimulated with 1 μM ISO for 30 s, and cell lysates were subjected to immuno-isolation with anti-pS355/356 (GRK-pβ 2 AR) specific antibody (1st immunoprecipitate, IP). The remaining β 2 ARs in the supernatant were isolated with anti-FLAG antibody (2nd IP). Total β 2 AR were immuno-isolated directly with anti-FLAG antibody (total IP). The β 2 AR in total IP and sequential IPs were resolved with SDS-PAGE, and probed with anti-FLAG, anti-pS261/262 (PKA-pβ 2 AR), and anti-pS355/356 (GRK-pβ 2 AR) antibodies, respectively. Representative of three independent experiments. Molecular weight markers (in kDa) are indicated on the left. Error bars denote s.e.m.; multiplicity adjusted P values are computed by one-way ANOVA followed by Tukey’s test between indicated groups

    Article Snippet: Other antibodies used in the experiments include: anti-FLAG-M1 and biotinylated anti-FLAG-M2 (F3040 and F9291, Sigma, MO), biotinylated goat anti-mouse IgG, and goat anti-rabbit IgG (111–065–144 and 115–055–166, Jackson ImmunoResearch, PA).

    Techniques: Imaging, Staining, Expressing, Isolation, SDS Page, Molecular Weight

    Subpopulations of β 2 AR display distinct structural properties. a Schematic of SiMPull with phospho-specific antibody against FLAG-mYFP-β 2 AR and examples of fluorescent time traces depicting one- or two-step photobleaching of mYFP-β 2 ARs, which provides stoichiometric information of β 2 AR. The composition of β 2 AR was calculated based on photobleaching steps in different conditions described in b and c . b Cells are stimulated with different concentrations of ISO for 5 min, the total β 2 ARs were pulled down for photobleaching analysis, displaying a composition of ~40% dimers and ~60% monomers independent of agonist stimulation (mean ± s.d.; n = 10, 8, 10, and 8 independent experiments, respectively). c The PKA- and GRK-phosphorylated β 2 ARs were pulled down with phospho-specific antibodies against S261/262 (PKA-pβ 2 AR) and S355/356 (GRK-pβ 2 AR) with different concentrations of ISO as indicated. At 1 nM, ISO only induces PKA-phosphorylation of β 2 AR but not GRK-phosphorylation of β 2 AR; higher concentrations of ISO induce both PKA- and GRK-phosphorylation of β 2 ARs. PKA-phosphorylated β 2 ARs display a composition of nearly 70% dimers and 30% monomers upon stimulation with all tested concentration of ISO (mean ± s.d.; n = 6, 6, and 14 independent experiments, respectively), whereas GRK-phosphorylated β 2 ARs display a composition of 12% dimers and 88% monomers at high concentrations of ISO stimulation (mean ± s.d.; n = 12 and 21 independent experiments, respectively). N.A. not available

    Journal: Nature Communications

    Article Title: Functionally distinct and selectively phosphorylated GPCR subpopulations co-exist in a single cell

    doi: 10.1038/s41467-018-03459-7

    Figure Lengend Snippet: Subpopulations of β 2 AR display distinct structural properties. a Schematic of SiMPull with phospho-specific antibody against FLAG-mYFP-β 2 AR and examples of fluorescent time traces depicting one- or two-step photobleaching of mYFP-β 2 ARs, which provides stoichiometric information of β 2 AR. The composition of β 2 AR was calculated based on photobleaching steps in different conditions described in b and c . b Cells are stimulated with different concentrations of ISO for 5 min, the total β 2 ARs were pulled down for photobleaching analysis, displaying a composition of ~40% dimers and ~60% monomers independent of agonist stimulation (mean ± s.d.; n = 10, 8, 10, and 8 independent experiments, respectively). c The PKA- and GRK-phosphorylated β 2 ARs were pulled down with phospho-specific antibodies against S261/262 (PKA-pβ 2 AR) and S355/356 (GRK-pβ 2 AR) with different concentrations of ISO as indicated. At 1 nM, ISO only induces PKA-phosphorylation of β 2 AR but not GRK-phosphorylation of β 2 AR; higher concentrations of ISO induce both PKA- and GRK-phosphorylation of β 2 ARs. PKA-phosphorylated β 2 ARs display a composition of nearly 70% dimers and 30% monomers upon stimulation with all tested concentration of ISO (mean ± s.d.; n = 6, 6, and 14 independent experiments, respectively), whereas GRK-phosphorylated β 2 ARs display a composition of 12% dimers and 88% monomers at high concentrations of ISO stimulation (mean ± s.d.; n = 12 and 21 independent experiments, respectively). N.A. not available

    Article Snippet: Other antibodies used in the experiments include: anti-FLAG-M1 and biotinylated anti-FLAG-M2 (F3040 and F9291, Sigma, MO), biotinylated goat anti-mouse IgG, and goat anti-rabbit IgG (111–065–144 and 115–055–166, Jackson ImmunoResearch, PA).

    Techniques: Concentration Assay

    Subcellular localization of TIG3-FLAG fusion proteins. A Schematic of the fusion proteins in which the segments of the N-terminal hydrophilic region are fused to FLAG epitope. The TIG3 domains are described in the legend to Fig. 1A . B SCC-13 cells were electroporated with 3 µg of each TIG3-FLAG fusion protein and after 24 h the cells were fixed and co-stained with the centrosome marker, pericentrin (green), and Cy3-conjugated anti-FLAG (red). Each of these fusion proteins localize at the centrosome (yellow, arrows). Bars = 10 µm. C TIG3(102–125)-FLAG accumulates at the centrosome in A431 cells and normal foreskin keratinocytes (KERn). Cells were electroporated with plasmid encoding TIG3(102–125)-FLAG and then stained with antibodies as indicated above. Bars = 10 µM.

    Journal: The Journal of investigative dermatology

    Article Title: Pericentrosomal localization of the TIG3 tumor suppressor requires an N-terminal hydrophilic region motif

    doi: 10.1038/jid.2013.533

    Figure Lengend Snippet: Subcellular localization of TIG3-FLAG fusion proteins. A Schematic of the fusion proteins in which the segments of the N-terminal hydrophilic region are fused to FLAG epitope. The TIG3 domains are described in the legend to Fig. 1A . B SCC-13 cells were electroporated with 3 µg of each TIG3-FLAG fusion protein and after 24 h the cells were fixed and co-stained with the centrosome marker, pericentrin (green), and Cy3-conjugated anti-FLAG (red). Each of these fusion proteins localize at the centrosome (yellow, arrows). Bars = 10 µm. C TIG3(102–125)-FLAG accumulates at the centrosome in A431 cells and normal foreskin keratinocytes (KERn). Cells were electroporated with plasmid encoding TIG3(102–125)-FLAG and then stained with antibodies as indicated above. Bars = 10 µM.

    Article Snippet: Antibodies and Reagents Polyclonal rabbit anti-TIG3 production was as previously described ( ; ; ). β-actin (A1978) and FLAG-M2-Cy3 (A9594) antibodies were purchased from Sigma (St Louis, MO).

    Techniques: FLAG-tag, Staining, Marker, Plasmid Preparation