fluorescein antibody Abcam Search Results


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  • 90
    Abcam rabbit polyclonal anti fluorescein isothiocyanate fitc
    TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A . At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein <t>isothiocyanate</t> <t>(FITC)-labeled</t> RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with <t>polyclonal</t> anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B . The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). * P
    Rabbit Polyclonal Anti Fluorescein Isothiocyanate Fitc, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam antifluorescein antibody
    TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A . At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein <t>isothiocyanate</t> <t>(FITC)-labeled</t> RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with <t>polyclonal</t> anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B . The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). * P
    Antifluorescein Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Abcam fluorescein tagged secondary antibody
    TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A . At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein <t>isothiocyanate</t> <t>(FITC)-labeled</t> RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with <t>polyclonal</t> anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B . The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). * P
    Fluorescein Tagged Secondary Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 78/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Abcam anti fluorescein antibody biotin
    TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A . At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein <t>isothiocyanate</t> <t>(FITC)-labeled</t> RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with <t>polyclonal</t> anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B . The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). * P
    Anti Fluorescein Antibody Biotin, supplied by Abcam, used in various techniques. Bioz Stars score: 84/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam anti fitc secondary antibody
    Superior tumor accumulation and retention of PEGylated <t>aptamer</t> than that of antibody. (a) PEGylated aptamer was developed by attaching a 20 kDa <t>PEG-FITC</t> to the 3'-end and a biotin or a DY647 dye to the 5'- end of the DNA strand. (b) Particle size of PEGylated aptamer and antibody as determined by dynamic light scattering. (c) Determination of the equilibrium dissociation constants (K'd) of PEGylated aptamer to HT29 cells using flow cytometry by incubating cells at varying concentrations (1-200 nmol/L). (d) Binding and internalization of PEGylated aptamer to HT29 cells which were incubated with 100 nM EpCAM aptamer at 37 °C for 30 min, followed by washing and confocal microscopy imaging. Scale bar = 10 μm. (e) Live animal imaging of antibodies and aptamers. NOD-SCID mice bearing HT29 tumor (150 mm 3 ) received a single intravenous injection of 0.75 nmol of control PEGylated aptamer, PEGylated aptamer and antibody followed by live animal imaging at the indicated time points. (f) The fluorescence-time curve of PEGylated aptamer in tumors as in (e) was determined by Living Imaging Software v2.50 (Xenogen) with the units of photons/s/cm 2 /sr. Log-scale heat map (at the right) of photon flux applies to all panels. Data are means ± SEM, n=3. RFI: relative fluorescence intensity.
    Anti Fitc Secondary Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Abcam secondary fluorescein conjugated antibodies
    Superior tumor accumulation and retention of PEGylated <t>aptamer</t> than that of antibody. (a) PEGylated aptamer was developed by attaching a 20 kDa <t>PEG-FITC</t> to the 3'-end and a biotin or a DY647 dye to the 5'- end of the DNA strand. (b) Particle size of PEGylated aptamer and antibody as determined by dynamic light scattering. (c) Determination of the equilibrium dissociation constants (K'd) of PEGylated aptamer to HT29 cells using flow cytometry by incubating cells at varying concentrations (1-200 nmol/L). (d) Binding and internalization of PEGylated aptamer to HT29 cells which were incubated with 100 nM EpCAM aptamer at 37 °C for 30 min, followed by washing and confocal microscopy imaging. Scale bar = 10 μm. (e) Live animal imaging of antibodies and aptamers. NOD-SCID mice bearing HT29 tumor (150 mm 3 ) received a single intravenous injection of 0.75 nmol of control PEGylated aptamer, PEGylated aptamer and antibody followed by live animal imaging at the indicated time points. (f) The fluorescence-time curve of PEGylated aptamer in tumors as in (e) was determined by Living Imaging Software v2.50 (Xenogen) with the units of photons/s/cm 2 /sr. Log-scale heat map (at the right) of photon flux applies to all panels. Data are means ± SEM, n=3. RFI: relative fluorescence intensity.
    Secondary Fluorescein Conjugated Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Abcam anti mouse fluorescein
    Superior tumor accumulation and retention of PEGylated <t>aptamer</t> than that of antibody. (a) PEGylated aptamer was developed by attaching a 20 kDa <t>PEG-FITC</t> to the 3'-end and a biotin or a DY647 dye to the 5'- end of the DNA strand. (b) Particle size of PEGylated aptamer and antibody as determined by dynamic light scattering. (c) Determination of the equilibrium dissociation constants (K'd) of PEGylated aptamer to HT29 cells using flow cytometry by incubating cells at varying concentrations (1-200 nmol/L). (d) Binding and internalization of PEGylated aptamer to HT29 cells which were incubated with 100 nM EpCAM aptamer at 37 °C for 30 min, followed by washing and confocal microscopy imaging. Scale bar = 10 μm. (e) Live animal imaging of antibodies and aptamers. NOD-SCID mice bearing HT29 tumor (150 mm 3 ) received a single intravenous injection of 0.75 nmol of control PEGylated aptamer, PEGylated aptamer and antibody followed by live animal imaging at the indicated time points. (f) The fluorescence-time curve of PEGylated aptamer in tumors as in (e) was determined by Living Imaging Software v2.50 (Xenogen) with the units of photons/s/cm 2 /sr. Log-scale heat map (at the right) of photon flux applies to all panels. Data are means ± SEM, n=3. RFI: relative fluorescence intensity.
    Anti Mouse Fluorescein, supplied by Abcam, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Abcam anti brdu fluorescein
    Superior tumor accumulation and retention of PEGylated <t>aptamer</t> than that of antibody. (a) PEGylated aptamer was developed by attaching a 20 kDa <t>PEG-FITC</t> to the 3'-end and a biotin or a DY647 dye to the 5'- end of the DNA strand. (b) Particle size of PEGylated aptamer and antibody as determined by dynamic light scattering. (c) Determination of the equilibrium dissociation constants (K'd) of PEGylated aptamer to HT29 cells using flow cytometry by incubating cells at varying concentrations (1-200 nmol/L). (d) Binding and internalization of PEGylated aptamer to HT29 cells which were incubated with 100 nM EpCAM aptamer at 37 °C for 30 min, followed by washing and confocal microscopy imaging. Scale bar = 10 μm. (e) Live animal imaging of antibodies and aptamers. NOD-SCID mice bearing HT29 tumor (150 mm 3 ) received a single intravenous injection of 0.75 nmol of control PEGylated aptamer, PEGylated aptamer and antibody followed by live animal imaging at the indicated time points. (f) The fluorescence-time curve of PEGylated aptamer in tumors as in (e) was determined by Living Imaging Software v2.50 (Xenogen) with the units of photons/s/cm 2 /sr. Log-scale heat map (at the right) of photon flux applies to all panels. Data are means ± SEM, n=3. RFI: relative fluorescence intensity.
    Anti Brdu Fluorescein, supplied by Abcam, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Abcam abcam α gfp antibody
    Transfection of Saos-2 cells with <t>GFP-RB,</t> <t>-p107,</t> and -p130 constructs results in expression of functional GFP-tagged pocket proteins exhibiting similar protein dynamics. A, eGFP was fused to the N terminus of wild-type RB, p107, and p130. A and B domains
    Abcam α Gfp Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 79/100, based on 572 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    Abcam fluorescein conjugated anti his6 antibody
    Transfection of Saos-2 cells with <t>GFP-RB,</t> <t>-p107,</t> and -p130 constructs results in expression of functional GFP-tagged pocket proteins exhibiting similar protein dynamics. A, eGFP was fused to the N terminus of wild-type RB, p107, and p130. A and B domains
    Fluorescein Conjugated Anti His6 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 83/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Abcam fluorescein isothiocyanate
    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein <t>isothiocyanate</t> conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.
    Fluorescein Isothiocyanate, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Abcam anti mouse fitc abcam
    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein <t>isothiocyanate</t> conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.
    Anti Mouse Fitc Abcam, supplied by Abcam, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Abcam anti fluorescein antibody conjugated to texas red
    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein <t>isothiocyanate</t> conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.
    Anti Fluorescein Antibody Conjugated To Texas Red, supplied by Abcam, used in various techniques. Bioz Stars score: 80/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Abcam fitc 8 oxodg abcam antibody
    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein <t>isothiocyanate</t> conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.
    Fitc 8 Oxodg Abcam Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 87/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam fluorescein isothiocyanate conjugated secondary antibody
    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein <t>isothiocyanate</t> conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.
    Fluorescein Isothiocyanate Conjugated Secondary Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Abcam fluorescein isothiocyanate conjugated vwf antibody
    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein <t>isothiocyanate</t> conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.
    Fluorescein Isothiocyanate Conjugated Vwf Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Abcam anti unc5b abcam fitc
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
    Anti Unc5b Abcam Fitc, supplied by Abcam, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Abcam fluorescein isothiocyanate conjugated h3k9me2 specific antibody
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
    Fluorescein Isothiocyanate Conjugated H3k9me2 Specific Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Abcam fluorescein labeled secondary antibodies
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
    Fluorescein Labeled Secondary Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam antihuman tlr 3 fluorescein isothiocynate fitc antibody
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
    Antihuman Tlr 3 Fluorescein Isothiocynate Fitc Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Abcam fluorescein labeled anti il 18r1
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
    Fluorescein Labeled Anti Il 18r1, supplied by Abcam, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Abcam fluorescein isothiocyanate anti chicken
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
    Fluorescein Isothiocyanate Anti Chicken, supplied by Abcam, used in various techniques. Bioz Stars score: 79/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti mouse fluorescein conjugated rabbit antibody
    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
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    Transendothelial neutrophil migration is inhibited by anti- <t>UNC5B</t> antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of <t>FITC</t> Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P
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    Abcam anti fitc antibody
    Interaction between TIPRL and eIF2α increases phosphorylation of eIF2α. a IP assays of cell lysates of A549 cells with anti-TIPRL antibody (upper panel) and cell lysates of 293 T cells with anti-HA antibody after transfection with gradual concentration of HA-tagged TIPRL (lower panel). b IP assays with anti-TIPRL antibody in A549 cells incubated in EBSS for 2 h. c Immunocytochemistry data of A549 cells where TIPRL and eIF2α proteins were double-immunostained and treated with goat anti-rabbit mouse <t>IgG-FITC</t> (green) and bovine anti-mouse IgG-Texas Red (red). Relative intensity of interaction were measured. d GST-pull down assay (left panel) and IP assay (right panel) with anti-HA antibody of 293 T cells co-transfected with HA-TIPRL along with GST-Mock, GST-eIF2α, GST-eIF2β, and GST-eIF2γ, respectively. e GST-pull down assay of 293T cells co-transfected with GST-eIF2α and TIPRL deletion mutants (D1–D6) (upper panel); six deletion fragments of TIPRL were sub-cloned into the pCGN-HA vector (lower panel). f IP assay of A549 cells mixed with TIPRL-mimic peptides which represent the regions of TIPRL spanning the amino acids 86–100 or 174–188. g In vitro kinase assays of phospho-eIF2α using recombinant eIF2α, gradual concentration of TIPRL, and <t>GCN2</t> proteins. Phosphorylation level of eIF2α was detected by Western blots, and recombinant proteins were stained using Ponceau S (left panel). Relative intensity of phospho-eIF2α were measured (right panel). h In vitro kinase assays of phospho-eIF2α using recombinant eIF2α, GCN2, and TIPRL proteins with incubation of 100 and 200 µM of TIPRL-mimic peptides. Phosphorylation of eIF2α and transfection of peptide were detected by Western blots, and recombinant proteins were stained using Ponceau S (left panel). Relative intensity of phospho-eIF2α were measured (right panel). Depicted western blots are representatives from 2–3 independent experiment. All quantitative bar data are mean ± SEM. p -value was calculated by t -test. * P
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    Abcam fluorescein isothiocyanate fitc conjugated secondary antibody
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Abcam fluorescein conjugated rabbit polyclonal anti c1q antibody
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Abcam fluorescein conjugated goat anti myc
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Abcam fluorescein isothiocyanate conjugated anti dr5 antibody
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Abcam fluorescein isothiocyanate tritc labeled secondary antibody
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Abcam fluorescein isothiocyanate fitc conjugated klf4 antibody
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Abcam donkey anti goat antibody fluorescein isothiocyanate
    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein <t>isothiocyanate</t> <t>(FITC)-conjugated</t> secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.
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    Image Search Results


    TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A . At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein isothiocyanate (FITC)-labeled RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with polyclonal anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B . The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). * P

    Journal: Molecular Brain

    Article Title: The long non-coding RNA nuclear-enriched abundant transcript 1_2 induces paraspeckle formation in the motor neuron during the early phase of amyotrophic lateral sclerosis

    doi: 10.1186/1756-6606-6-31

    Figure Lengend Snippet: TDP-43 and FUS/TLS aggregate at nuclear NEAT1_2 foci. A . At 48 hours after transfection with wild-type (WT) and 35-kDa TDP-43 fragment and WT FUS/TLS with the V5 tag at the C terminus, HeLa cells were fixed with 4% paraformaldehyde, hybridized with fluorescein isothiocyanate (FITC)-labeled RNA probe against NEAT1_2 long non-coding RNA (lncRNA), and double-immunostained with polyclonal anti-FITC and monoclonal anti-V5 antibodies. Schematic diagram (right) represents the TDP-43 isoforms and FUS/TLS. NLS, Nuclear localization signal; RRM, RNA recognition motif; C-term, C-terminal domain; GR, glycine-rich motif; NES, nuclear export signal; RGG, arginine-glycine-glycine motif ; ZF, zinc-finger motif. Dotted lines represent the outline of the nucleus. Scale bars, 10μm. B . The Y-axis shows what percent NEAT1_2 foci is overlapped by TDP-43 nuclear aggregates in (A). The 26-kDa TDP-43 fragment hardly accumulates in nuclei and lacks affinity for NEAT1_2 foci. Data represent mean ± s.d. (n = 50 for each transfection of TDP-43 isoforms). * P

    Article Snippet: Rabbit polyclonal anti-fluorescein isothiocyanate (FITC) (1:500, ab19491), mouse monoclonal anti-digoxigenin (DIG) IgG1 (1:500, H8, ab420), and mouse monoclonal anti-coilin IgG2b (1:500, IH10, ab87913) were from Abcam (Cambridge, MA).

    Techniques: Transfection, Labeling

    Superior tumor accumulation and retention of PEGylated aptamer than that of antibody. (a) PEGylated aptamer was developed by attaching a 20 kDa PEG-FITC to the 3'-end and a biotin or a DY647 dye to the 5'- end of the DNA strand. (b) Particle size of PEGylated aptamer and antibody as determined by dynamic light scattering. (c) Determination of the equilibrium dissociation constants (K'd) of PEGylated aptamer to HT29 cells using flow cytometry by incubating cells at varying concentrations (1-200 nmol/L). (d) Binding and internalization of PEGylated aptamer to HT29 cells which were incubated with 100 nM EpCAM aptamer at 37 °C for 30 min, followed by washing and confocal microscopy imaging. Scale bar = 10 μm. (e) Live animal imaging of antibodies and aptamers. NOD-SCID mice bearing HT29 tumor (150 mm 3 ) received a single intravenous injection of 0.75 nmol of control PEGylated aptamer, PEGylated aptamer and antibody followed by live animal imaging at the indicated time points. (f) The fluorescence-time curve of PEGylated aptamer in tumors as in (e) was determined by Living Imaging Software v2.50 (Xenogen) with the units of photons/s/cm 2 /sr. Log-scale heat map (at the right) of photon flux applies to all panels. Data are means ± SEM, n=3. RFI: relative fluorescence intensity.

    Journal: Theranostics

    Article Title: Superior Performance of Aptamer in Tumor Penetration over Antibody: Implication of Aptamer-Based Theranostics in Solid Tumors

    doi: 10.7150/thno.11711

    Figure Lengend Snippet: Superior tumor accumulation and retention of PEGylated aptamer than that of antibody. (a) PEGylated aptamer was developed by attaching a 20 kDa PEG-FITC to the 3'-end and a biotin or a DY647 dye to the 5'- end of the DNA strand. (b) Particle size of PEGylated aptamer and antibody as determined by dynamic light scattering. (c) Determination of the equilibrium dissociation constants (K'd) of PEGylated aptamer to HT29 cells using flow cytometry by incubating cells at varying concentrations (1-200 nmol/L). (d) Binding and internalization of PEGylated aptamer to HT29 cells which were incubated with 100 nM EpCAM aptamer at 37 °C for 30 min, followed by washing and confocal microscopy imaging. Scale bar = 10 μm. (e) Live animal imaging of antibodies and aptamers. NOD-SCID mice bearing HT29 tumor (150 mm 3 ) received a single intravenous injection of 0.75 nmol of control PEGylated aptamer, PEGylated aptamer and antibody followed by live animal imaging at the indicated time points. (f) The fluorescence-time curve of PEGylated aptamer in tumors as in (e) was determined by Living Imaging Software v2.50 (Xenogen) with the units of photons/s/cm 2 /sr. Log-scale heat map (at the right) of photon flux applies to all panels. Data are means ± SEM, n=3. RFI: relative fluorescence intensity.

    Article Snippet: For the detection of FITC-labeled antibody or FITC-labeled-aptamer, the sections were incubated with anti-FITC secondary antibody (Abcam, Cat #AB6656, 1: 100 dilution) for 2 h at room temperature.

    Techniques: Flow Cytometry, Cytometry, Binding Assay, Incubation, Confocal Microscopy, Imaging, Mouse Assay, Injection, Fluorescence, Software

    Transfection of Saos-2 cells with GFP-RB, -p107, and -p130 constructs results in expression of functional GFP-tagged pocket proteins exhibiting similar protein dynamics. A, eGFP was fused to the N terminus of wild-type RB, p107, and p130. A and B domains

    Journal: The Journal of Biological Chemistry

    Article Title: Retinoblastoma/p107/p130 Pocket Proteins

    doi: 10.1074/jbc.M808740200

    Figure Lengend Snippet: Transfection of Saos-2 cells with GFP-RB, -p107, and -p130 constructs results in expression of functional GFP-tagged pocket proteins exhibiting similar protein dynamics. A, eGFP was fused to the N terminus of wild-type RB, p107, and p130. A and B domains

    Article Snippet: As with p107 and p130, the GFP antibody was able to immunoprecipitate chromatin-associated GFP-RB specifically at E2F target promoters.

    Techniques: Transfection, Construct, Expressing, Functional Assay

    Antibodies recognizing GFP can be used to detect p107, p130, and RB at target gene promoters. A, chromatin was isolated from Saos-2 cells 48 h following transfection with GFP-p107. ChIP assays were performed using three commercially available GFP antibodies

    Journal: The Journal of Biological Chemistry

    Article Title: Retinoblastoma/p107/p130 Pocket Proteins

    doi: 10.1074/jbc.M808740200

    Figure Lengend Snippet: Antibodies recognizing GFP can be used to detect p107, p130, and RB at target gene promoters. A, chromatin was isolated from Saos-2 cells 48 h following transfection with GFP-p107. ChIP assays were performed using three commercially available GFP antibodies

    Article Snippet: As with p107 and p130, the GFP antibody was able to immunoprecipitate chromatin-associated GFP-RB specifically at E2F target promoters.

    Techniques: Isolation, Transfection, Chromatin Immunoprecipitation

    Degradation of Os WRKY11 protein by the UPS. a Rice ( Oryza sativa ) protoplasts were transformed with 35S:: OsWRKY11-YFP . After 4 h, protoplasts were treated with 100 μM MG132 and buffer for 1 day. Fluorescence was observed using a confocal laser scanning microscope. Scale bars: 30 μm. NT: non-treated control. b Immunoblot analysis of total protoplast proteins in the absence (−) or presence (+) of 100 μM MG132. Blots were probed with green fluorescent protein (GFP) antibody. The PAT antibody was used as a control for transformation efficiency

    Journal: Rice

    Article Title: Rice WRKY11 Plays a Role in Pathogen Defense and Drought Tolerance

    doi: 10.1186/s12284-018-0199-0

    Figure Lengend Snippet: Degradation of Os WRKY11 protein by the UPS. a Rice ( Oryza sativa ) protoplasts were transformed with 35S:: OsWRKY11-YFP . After 4 h, protoplasts were treated with 100 μM MG132 and buffer for 1 day. Fluorescence was observed using a confocal laser scanning microscope. Scale bars: 30 μm. NT: non-treated control. b Immunoblot analysis of total protoplast proteins in the absence (−) or presence (+) of 100 μM MG132. Blots were probed with green fluorescent protein (GFP) antibody. The PAT antibody was used as a control for transformation efficiency

    Article Snippet: Immunoblot analysis was performed with GFP antibody (Abcam; code: ab6556).

    Techniques: Transformation Assay, Fluorescence, Laser-Scanning Microscopy

    Effect of αvβ5 and αvβ3 integrin overexpression and knock-down on FA formation. hCFs were transfected with ( A ) GFP-tagged constructs to express αvβ5 and αvβ3 integrins, ( B ) shRNA targeting

    Journal: Cardiovascular Research

    Article Title: Integrins αvβ5 and αvβ3 promote latent TGF-β1 activation by human cardiac fibroblast contraction

    doi: 10.1093/cvr/cvu053

    Figure Lengend Snippet: Effect of αvβ5 and αvβ3 integrin overexpression and knock-down on FA formation. hCFs were transfected with ( A ) GFP-tagged constructs to express αvβ5 and αvβ3 integrins, ( B ) shRNA targeting

    Article Snippet: Primary antibodies directed against α-SMA (α-SM1, Giulio Gabbiani, University of Geneva, Switzerland), α-sarcomeric actin (A2172, Sigma-Aldrich), CD31 (ab28364, Abcam, Cambridge, MA, USA), desmin (M076029, Dako), GFP (ab290, Abcam), integrin β3 (for hCF: Axum-4, Dr Dean Sheppard and for porcine tissues: MAB1974, Millipore, Etobicoke, ON, Canada), integrin β5 (ab15459, Abcam), vimentin (M0725, Dako), and vinculin (V9264, Sigma-Aldrich) were used.

    Techniques: Over Expression, Transfection, Construct, shRNA

    Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein isothiocyanate conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.

    Journal: Molecular Medicine Reports

    Article Title: Role of glycoprotein 78 and cidec in hepatic steatosis

    doi: 10.3892/mmr.2017.6834

    Figure Lengend Snippet: Association between gp78 and cidec in hepatic steatosis. (A) Nuclei were stained with DAPI, (a) gp78 was visualized (fluorescein isothiocyanate conjugated secondary antibody; red staining). (b) Cidec was visualized (tetraethyl rhodamine isothiocyanate conjugated secondary antibody; green staining); (c) Colocalization of gp78 and cidec (yellow) visualized on the surface of lipid droplets using immunofluorescent microscopy and identified by coimmunoprecipitation (scale bar=15 µm). (B) Gp78 and Cidec direct interaction detected by coimmunoprecipitation assay. gp78, glycoprotein; cidec, cell death-inducing DFFA-like effector c.

    Article Snippet: The cells were then incubated with primary antibodies specific for gp78 (ab54787; dilutions 1:1,000, Abcam) or cidec (ab213693; dilutions 1:1,000, Abcam) overnight at 4°C, followed by incubation with secondary antibodies conjugated with fluorescein isothiocyanate for gp78 (ab6785; dilutions 1:3,000, Abcam) and with tetraethyl rhodamine isothiocyanate for cidec (ab6718; 1:3,000, Abcam) for 1 h at 37°C.

    Techniques: Staining, Microscopy, Co-Immunoprecipitation Assay

    Transendothelial neutrophil migration is inhibited by anti- UNC5B antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of FITC Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P

    Journal: PLoS ONE

    Article Title: The Uncoordinated-5 Homolog B (UNC5B) Receptor Increases Myocardial Ischemia-Reperfusion Injury

    doi: 10.1371/journal.pone.0069477

    Figure Lengend Snippet: Transendothelial neutrophil migration is inhibited by anti- UNC5B antibody. A ) PMN transmigration studies were performed in transwells using fMLP as chemoattractant in the lower compartment. PMNs were pre-incubated with either anti-UNC5B antibody or IgG control prior to transmigration. Number of transmigrated PMNs was assessed by MPO measurement. B ) Passive flux of FITC Dextran across HMEC endothelium monolayers. Flux was detected by measurement of passed FITC Dextran in the basal compartment (n = 6 per condition; * P

    Article Snippet: As detection antibodies we used: anti-UNC5B (Abcam) FITC labeled, PE-mouse anti-human for CD3+ CD19; CD14 and CD15 detection and PerCP-mouse anti-human for CD45 detection (all BD Bioscience).

    Techniques: Migration, Transmigration Assay, Incubation

    Interaction between TIPRL and eIF2α increases phosphorylation of eIF2α. a IP assays of cell lysates of A549 cells with anti-TIPRL antibody (upper panel) and cell lysates of 293 T cells with anti-HA antibody after transfection with gradual concentration of HA-tagged TIPRL (lower panel). b IP assays with anti-TIPRL antibody in A549 cells incubated in EBSS for 2 h. c Immunocytochemistry data of A549 cells where TIPRL and eIF2α proteins were double-immunostained and treated with goat anti-rabbit mouse IgG-FITC (green) and bovine anti-mouse IgG-Texas Red (red). Relative intensity of interaction were measured. d GST-pull down assay (left panel) and IP assay (right panel) with anti-HA antibody of 293 T cells co-transfected with HA-TIPRL along with GST-Mock, GST-eIF2α, GST-eIF2β, and GST-eIF2γ, respectively. e GST-pull down assay of 293T cells co-transfected with GST-eIF2α and TIPRL deletion mutants (D1–D6) (upper panel); six deletion fragments of TIPRL were sub-cloned into the pCGN-HA vector (lower panel). f IP assay of A549 cells mixed with TIPRL-mimic peptides which represent the regions of TIPRL spanning the amino acids 86–100 or 174–188. g In vitro kinase assays of phospho-eIF2α using recombinant eIF2α, gradual concentration of TIPRL, and GCN2 proteins. Phosphorylation level of eIF2α was detected by Western blots, and recombinant proteins were stained using Ponceau S (left panel). Relative intensity of phospho-eIF2α were measured (right panel). h In vitro kinase assays of phospho-eIF2α using recombinant eIF2α, GCN2, and TIPRL proteins with incubation of 100 and 200 µM of TIPRL-mimic peptides. Phosphorylation of eIF2α and transfection of peptide were detected by Western blots, and recombinant proteins were stained using Ponceau S (left panel). Relative intensity of phospho-eIF2α were measured (right panel). Depicted western blots are representatives from 2–3 independent experiment. All quantitative bar data are mean ± SEM. p -value was calculated by t -test. * P

    Journal: Cell Death & Disease

    Article Title: TIPRL potentiates survival of lung cancer by inducing autophagy through the eIF2α-ATF4 pathway

    doi: 10.1038/s41419-019-2190-0

    Figure Lengend Snippet: Interaction between TIPRL and eIF2α increases phosphorylation of eIF2α. a IP assays of cell lysates of A549 cells with anti-TIPRL antibody (upper panel) and cell lysates of 293 T cells with anti-HA antibody after transfection with gradual concentration of HA-tagged TIPRL (lower panel). b IP assays with anti-TIPRL antibody in A549 cells incubated in EBSS for 2 h. c Immunocytochemistry data of A549 cells where TIPRL and eIF2α proteins were double-immunostained and treated with goat anti-rabbit mouse IgG-FITC (green) and bovine anti-mouse IgG-Texas Red (red). Relative intensity of interaction were measured. d GST-pull down assay (left panel) and IP assay (right panel) with anti-HA antibody of 293 T cells co-transfected with HA-TIPRL along with GST-Mock, GST-eIF2α, GST-eIF2β, and GST-eIF2γ, respectively. e GST-pull down assay of 293T cells co-transfected with GST-eIF2α and TIPRL deletion mutants (D1–D6) (upper panel); six deletion fragments of TIPRL were sub-cloned into the pCGN-HA vector (lower panel). f IP assay of A549 cells mixed with TIPRL-mimic peptides which represent the regions of TIPRL spanning the amino acids 86–100 or 174–188. g In vitro kinase assays of phospho-eIF2α using recombinant eIF2α, gradual concentration of TIPRL, and GCN2 proteins. Phosphorylation level of eIF2α was detected by Western blots, and recombinant proteins were stained using Ponceau S (left panel). Relative intensity of phospho-eIF2α were measured (right panel). h In vitro kinase assays of phospho-eIF2α using recombinant eIF2α, GCN2, and TIPRL proteins with incubation of 100 and 200 µM of TIPRL-mimic peptides. Phosphorylation of eIF2α and transfection of peptide were detected by Western blots, and recombinant proteins were stained using Ponceau S (left panel). Relative intensity of phospho-eIF2α were measured (right panel). Depicted western blots are representatives from 2–3 independent experiment. All quantitative bar data are mean ± SEM. p -value was calculated by t -test. * P

    Article Snippet: Antibody and western blotting The anti-LC3B antibody (L7543) was purchased from Sigma-Aldrich (MO, USA); HA (LF-MA0048) and GAPDH (LF-PA0212) antibodies were purchased from Ab frontier (Seoul, Republic of Korea); phospho-GCN2(T899) (ab75836), HSP60 (ab46798), Cytochrome c (ab76107), FITC (ab19224) antibodies were purchased from abcam (MA, USA); P62 (610832), HIF1α (610959) antibodies were purchased from BD bioscience (CA, USA); TIPRL antibody (a300-663a) was purchased from bethyl (TX, USA); Bip (3183), phosphor-mTOR(s2448) (5536), mTOR (2983), p-p70 s6k(T389) (9234), p-4e-bp1(T37/46) (2855), p-eIF2α(S51) (9721), eIF2α (9722), PERK (3192), MKK7 (4172), Caspase-8 (9746), Caspase-3 (9665), PARP (9542) antibodies were purchased from Cell Signaling Technology (MA, USA); β-actin (sc-47778), ATG7 (sc-33211), GCN2 (sc374609), ATF4 (sc-200), phospho-PERK(T981) (sc-32577), GST (sc-138), PP1 (sc-7482), GADD34 (sc-8327), normal mouse IgG (sc-2025), normal rabbit IgG (sc-2027), TOM20 (sc-17764), Nbr1 (sc-130380), PP2A-Cα/β (sc-6110), Goat-anti Rabbit IgG-HRP (sc-2004), Goat-anti Mouse IgG-HRP (sc-2031), Donkey-anti Goat IgG-HRP (sc-2033) antibodies were purchased from Santa Cruz (CA, USA).

    Techniques: Transfection, Concentration Assay, Incubation, Immunocytochemistry, Pull Down Assay, Clone Assay, Plasmid Preparation, In Vitro, Recombinant, Western Blot, Staining

    PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein isothiocyanate (FITC)-conjugated secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.

    Journal: PLoS ONE

    Article Title: The Natural Human IgM Antibody PAT-SM6 Induces Apoptosis in Primary Human Multiple Myeloma Cells by Targeting Heat Shock Protein GRP78

    doi: 10.1371/journal.pone.0063414

    Figure Lengend Snippet: PAT-SM6 specifically binds to MM cell lines and primary human MM samples. A, INA-6, NCI H929, MM1.S, OPM-2 and U266 MM cell lines or CD138-purified patient MM cells were stained with PAT-SM6 (5 µg/mL, open histogram) or isotype control (gray histogram) followed by fluorescein isothiocyanate (FITC)-conjugated secondary antibodies and assessed by flow cytometry. Both, cell lines as well as primary MM cells displayed specific PAT-SM6 binding. B, Immunohistochemistry with PAT-SM6, CD138-positive control antibody or isotype on bone marrow biopsies of MM patients. PAT-SM6 specifically stained MM cells with a homogenous binding pattern. In samples without MM infiltration no specific binding was detected. Images were captured using a Leica DM BL microscope, the Leica ICC HD digital camera and the Leica LAS EZ V2.1.0 software.

    Article Snippet: The expression of GRP78 on MM cells was assessed using anti-GRP78 IgG (rabbit), as well as PAT-SM6 followed by fluorescein isothiocyanate (FITC)-conjugated secondary antibodies (Abcam or Dako).

    Techniques: Purification, Staining, Flow Cytometry, Cytometry, Binding Assay, Immunohistochemistry, Positive Control, Microscopy, Immunocytochemistry, Software