anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eea1
    Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eea1
    Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eea1
    Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti eea1
    Rabbit Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc eea1
    Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit monoclonal early endosome antigen 1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal early endosome antigen 1
    Effects of GM1 treatment on PDGFR activation. (a) Western blot analysis and quantification of PDGFR- β activation. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1, as compared to hTSCs differentiated in GM1-free osteogenic medium (O.D.). Total proteins were extracted and analyzed with anti-phosphorylated-PDGFR- β (Tyr 751) antibody (green) and anti-PDGFR- β (28E1) antibody (red). <t>EEA1</t> expression was used as internal control. Data are means ± SD of four different experiments. (b, c) Gene expression analysis of the osteogenic markers ALP and osteocalcin by real-time PCR. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1 or 10 ng/ml PDGF-BB or with both 100 μ M GM1 and 10 ng/ml PDGF-BB. The results were compared to hTSCs differentiated in free osteogenic medium (O.D.). Ribosomal protein S14 gene was used as housekeeper. All data are means ± SD of three different experiments. The statistical analysis was determined by Student's t-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.
    Rabbit Monoclonal Early Endosome Antigen 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "GM1 Ganglioside Promotes Osteogenic Differentiation of Human Tendon Stem Cells"

    Article Title: GM1 Ganglioside Promotes Osteogenic Differentiation of Human Tendon Stem Cells

    Journal: Stem Cells International

    doi: 10.1155/2018/4706943

    Effects of GM1 treatment on PDGFR activation. (a) Western blot analysis and quantification of PDGFR- β activation. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1, as compared to hTSCs differentiated in GM1-free osteogenic medium (O.D.). Total proteins were extracted and analyzed with anti-phosphorylated-PDGFR- β (Tyr 751) antibody (green) and anti-PDGFR- β (28E1) antibody (red). EEA1 expression was used as internal control. Data are means ± SD of four different experiments. (b, c) Gene expression analysis of the osteogenic markers ALP and osteocalcin by real-time PCR. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1 or 10 ng/ml PDGF-BB or with both 100 μ M GM1 and 10 ng/ml PDGF-BB. The results were compared to hTSCs differentiated in free osteogenic medium (O.D.). Ribosomal protein S14 gene was used as housekeeper. All data are means ± SD of three different experiments. The statistical analysis was determined by Student's t-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.
    Figure Legend Snippet: Effects of GM1 treatment on PDGFR activation. (a) Western blot analysis and quantification of PDGFR- β activation. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1, as compared to hTSCs differentiated in GM1-free osteogenic medium (O.D.). Total proteins were extracted and analyzed with anti-phosphorylated-PDGFR- β (Tyr 751) antibody (green) and anti-PDGFR- β (28E1) antibody (red). EEA1 expression was used as internal control. Data are means ± SD of four different experiments. (b, c) Gene expression analysis of the osteogenic markers ALP and osteocalcin by real-time PCR. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1 or 10 ng/ml PDGF-BB or with both 100 μ M GM1 and 10 ng/ml PDGF-BB. The results were compared to hTSCs differentiated in free osteogenic medium (O.D.). Ribosomal protein S14 gene was used as housekeeper. All data are means ± SD of three different experiments. The statistical analysis was determined by Student's t-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Techniques Used: Activation Assay, Western Blot, Expressing, Real-time Polymerase Chain Reaction

    rabbit anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti eea1
    <t>EEA1</t> undergoes mono-ubiquitination in cells. A , Cell extracts prepared from HEK293 cells transfected with HA-tagged ubiquitin- (HA-Ub) and EEA1-FLAG-expressing plasmids (+) or a control empty vector (−) were subject to immunoprecipitation with FLAG antibodies. The precipitated materials were analyzed by immunoblotting (IB) with the indicated antibodies. The bracket indicates ubiquitinated EEA1 species. B, As in A , except that cells expressing HA-Ub were used and that immunoprecipitation was performed with <t>anti-EEA1</t> antibodies. C, As in B , except that cells expressing the indicated Ub variants were used.
    Rabbit Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Monoubiquitination of EEA1 regulates endosome fusion and trafficking"

    Article Title: Monoubiquitination of EEA1 regulates endosome fusion and trafficking

    Journal: Cell & Bioscience

    doi: 10.1186/2045-3701-3-24

    EEA1 undergoes mono-ubiquitination in cells. A , Cell extracts prepared from HEK293 cells transfected with HA-tagged ubiquitin- (HA-Ub) and EEA1-FLAG-expressing plasmids (+) or a control empty vector (−) were subject to immunoprecipitation with FLAG antibodies. The precipitated materials were analyzed by immunoblotting (IB) with the indicated antibodies. The bracket indicates ubiquitinated EEA1 species. B, As in A , except that cells expressing HA-Ub were used and that immunoprecipitation was performed with anti-EEA1 antibodies. C, As in B , except that cells expressing the indicated Ub variants were used.
    Figure Legend Snippet: EEA1 undergoes mono-ubiquitination in cells. A , Cell extracts prepared from HEK293 cells transfected with HA-tagged ubiquitin- (HA-Ub) and EEA1-FLAG-expressing plasmids (+) or a control empty vector (−) were subject to immunoprecipitation with FLAG antibodies. The precipitated materials were analyzed by immunoblotting (IB) with the indicated antibodies. The bracket indicates ubiquitinated EEA1 species. B, As in A , except that cells expressing HA-Ub were used and that immunoprecipitation was performed with anti-EEA1 antibodies. C, As in B , except that cells expressing the indicated Ub variants were used.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Western Blot

    EEA1 can interact with E2 to undergo E3-independent mono-ubiquitination. A , In vitro ubiquitination was performed with a post-nuclear extract isolated from cells expressing EEA1-FLAG in the presence of HA-tagged ubiquitin and ATP as indicated. The reactions were analyzed by dual color immunoblotting with the indicated antibodies. B , A Coomassie-blue stained gel showing the purified EEA1 (indicated by the arrow). The bracket indicates 3XFLAG peptide. C , Purified EEA1 was incubated with the indicated E2 ubiquitin conjugating enzymes together with E1, HA-Ub, and ATP. Where indicated, EEA1-FLAG was omitted as negative controls. The reactions were analyzed by immunoblotting with the HA (top panel) and EEA1 antibodies (bottom panel). D , As in A, except that the indicated ubiquitin variants were used. E , F , Interaction of EEA1 with Ube2A. E , Purified Ube2A was injected into a CM5 chip immobilized with EEA1-FLAG at the indicated concentrations. The interaction of Ube2A with EEA1 was monitored by Surface Plasmon Resonance. F , The interaction of EEA1-FLAG with Ube2A was plotted against the E2 concentrations.
    Figure Legend Snippet: EEA1 can interact with E2 to undergo E3-independent mono-ubiquitination. A , In vitro ubiquitination was performed with a post-nuclear extract isolated from cells expressing EEA1-FLAG in the presence of HA-tagged ubiquitin and ATP as indicated. The reactions were analyzed by dual color immunoblotting with the indicated antibodies. B , A Coomassie-blue stained gel showing the purified EEA1 (indicated by the arrow). The bracket indicates 3XFLAG peptide. C , Purified EEA1 was incubated with the indicated E2 ubiquitin conjugating enzymes together with E1, HA-Ub, and ATP. Where indicated, EEA1-FLAG was omitted as negative controls. The reactions were analyzed by immunoblotting with the HA (top panel) and EEA1 antibodies (bottom panel). D , As in A, except that the indicated ubiquitin variants were used. E , F , Interaction of EEA1 with Ube2A. E , Purified Ube2A was injected into a CM5 chip immobilized with EEA1-FLAG at the indicated concentrations. The interaction of Ube2A with EEA1 was monitored by Surface Plasmon Resonance. F , The interaction of EEA1-FLAG with Ube2A was plotted against the E2 concentrations.

    Techniques Used: In Vitro, Isolation, Expressing, Western Blot, Staining, Purification, Incubation, Injection, SPR Assay

    Ub-EEA1 expression induced vacuole-like endocytic structures. A , COS7 cells transfected with the indicated plasmids were immunostained with anti-EEA1 antibodies in red. Cells were counter-stained with DAPI to reveal the nuclei. B , Quantification of Ub-EEA1-induced giant vacuole phenotype. The percentage of cells with enlarged vacuole-like endosomes in either EEA1 or Ub-EEA1 expressing cells is indicated. Error bars, SD (n = 3). C , EEA1- or Ub-EEA1-expressing cells were fixed and analyzed by transmission electron microscopy. The arrow indicates a representative vacuole-like structure in Ub-EEA1-expressing cells. D , Extracts prepared from cells transfected with the indicated plasmids were analyzed by immunoblotting to determine the expression levels of the indicated proteins.
    Figure Legend Snippet: Ub-EEA1 expression induced vacuole-like endocytic structures. A , COS7 cells transfected with the indicated plasmids were immunostained with anti-EEA1 antibodies in red. Cells were counter-stained with DAPI to reveal the nuclei. B , Quantification of Ub-EEA1-induced giant vacuole phenotype. The percentage of cells with enlarged vacuole-like endosomes in either EEA1 or Ub-EEA1 expressing cells is indicated. Error bars, SD (n = 3). C , EEA1- or Ub-EEA1-expressing cells were fixed and analyzed by transmission electron microscopy. The arrow indicates a representative vacuole-like structure in Ub-EEA1-expressing cells. D , Extracts prepared from cells transfected with the indicated plasmids were analyzed by immunoblotting to determine the expression levels of the indicated proteins.

    Techniques Used: Expressing, Transfection, Staining, Transmission Assay, Electron Microscopy, Western Blot

    Expression of Ub-EEA1 alters Golgi morphology and induces fusion of lysosome with endosome. COS7 cells expressing Ub-EEA1 were fixed and immunostained with EEA1 antibody ( B , E , H , K ) together with antibodies against β-COP ( A ), PDI ( D ), Tom20 ( G ), or Lamp1 ( J ). Cells were also stained with DAPI in blue to reveal the nuclei. Merged two-color images are shown in C , F , I , L . Scale bars correspond to 20 μm.
    Figure Legend Snippet: Expression of Ub-EEA1 alters Golgi morphology and induces fusion of lysosome with endosome. COS7 cells expressing Ub-EEA1 were fixed and immunostained with EEA1 antibody ( B , E , H , K ) together with antibodies against β-COP ( A ), PDI ( D ), Tom20 ( G ), or Lamp1 ( J ). Cells were also stained with DAPI in blue to reveal the nuclei. Merged two-color images are shown in C , F , I , L . Scale bars correspond to 20 μm.

    Techniques Used: Expressing, Staining

    Expression of Ub-EEA1 inhibits transferrin uptake. COS7 cells transfected with EEA1- or Ub-EEA1-expressing plasmids were treated with Texas red-labeled transferrin on ice. After removal of unbound transferrin, cells were incubated in a transferrin-free medium at 37°C for the indicated time points. Cells were fixed and stained with anti-EEA1 antibody (green) and DAPI (blue). Note that the highlighted Ub-EEA1-expressing cells fail to take up transferrin compared to the surrounding untransfected cells.
    Figure Legend Snippet: Expression of Ub-EEA1 inhibits transferrin uptake. COS7 cells transfected with EEA1- or Ub-EEA1-expressing plasmids were treated with Texas red-labeled transferrin on ice. After removal of unbound transferrin, cells were incubated in a transferrin-free medium at 37°C for the indicated time points. Cells were fixed and stained with anti-EEA1 antibody (green) and DAPI (blue). Note that the highlighted Ub-EEA1-expressing cells fail to take up transferrin compared to the surrounding untransfected cells.

    Techniques Used: Expressing, Transfection, Labeling, Incubation, Staining

    Expression of Ub-EEA1 blocks the recycling of the transferrin receptor. COS7 cells transfected with the Ub-EEA1-expressing plasmid were fixed and immunostained with an antibody against transferrin receptor (TfR) ( A , red channel in C) and anti-EEA1 antibody ( B , green channel in C). Cells were also stained with DAPI in blue. Arrow heads label two untransfected cells that contain many transferrin receptor molecules in recycling vesicles. By contrast, the circled Ub-EEA1-expressing cells contain few TfR positive recycling vesicles. The scale bars correspond to 20 μm.
    Figure Legend Snippet: Expression of Ub-EEA1 blocks the recycling of the transferrin receptor. COS7 cells transfected with the Ub-EEA1-expressing plasmid were fixed and immunostained with an antibody against transferrin receptor (TfR) ( A , red channel in C) and anti-EEA1 antibody ( B , green channel in C). Cells were also stained with DAPI in blue. Arrow heads label two untransfected cells that contain many transferrin receptor molecules in recycling vesicles. By contrast, the circled Ub-EEA1-expressing cells contain few TfR positive recycling vesicles. The scale bars correspond to 20 μm.

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Staining

    rabbit anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti eea1
    TmTNF-dependent vesicular trafficking of humanized anti-TNFs and detection of cell surface anti-TNF peptides in dendritic cells. (A) Compartmentalization of anti-TNF: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and the cells ‘pulsed’ on ice using Alexa 488-conjugated anti-TNF (0 min, ice). The fluorescent label on the anti-TNF was ‘chased’ for up to 1 h at 37°C. Endosomes and lysosomes were stained with Alexa 647-conjuated <t>anti-EEA1</t> and anti-LAMP1, respectively. Nucleus was stained with DAPI. Images were acquired using confocal microscopy. (B) Identification of cell surface-associated anti-TNF peptides: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and incubated with either an anti-TNF mAb or a DVD-Ig containing one anti-TNF domain for 6 h. Cell surface displayed peptides were eluted under mild acidic conditions and the identity of anti-TNF peptides was obtained using LC-MS/MS. (C) Identification of HLA-DR-associated anti-TNF peptides: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and incubated with anti-TNF for up to 8 h. HLA-DR was immunoprecipitated and the identity of HLA-DR-associated anti-TNF peptides was obtained using LC-MS/MS (underlined: linker peptide sequence of anti-TNF DVD-Ig).
    Rabbit Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Transmembrane TNF-dependent uptake of anti-TNF antibodies"

    Article Title: Transmembrane TNF-dependent uptake of anti-TNF antibodies

    Journal: mAbs

    doi: 10.1080/19420862.2017.1304869

    TmTNF-dependent vesicular trafficking of humanized anti-TNFs and detection of cell surface anti-TNF peptides in dendritic cells. (A) Compartmentalization of anti-TNF: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and the cells ‘pulsed’ on ice using Alexa 488-conjugated anti-TNF (0 min, ice). The fluorescent label on the anti-TNF was ‘chased’ for up to 1 h at 37°C. Endosomes and lysosomes were stained with Alexa 647-conjuated anti-EEA1 and anti-LAMP1, respectively. Nucleus was stained with DAPI. Images were acquired using confocal microscopy. (B) Identification of cell surface-associated anti-TNF peptides: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and incubated with either an anti-TNF mAb or a DVD-Ig containing one anti-TNF domain for 6 h. Cell surface displayed peptides were eluted under mild acidic conditions and the identity of anti-TNF peptides was obtained using LC-MS/MS. (C) Identification of HLA-DR-associated anti-TNF peptides: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and incubated with anti-TNF for up to 8 h. HLA-DR was immunoprecipitated and the identity of HLA-DR-associated anti-TNF peptides was obtained using LC-MS/MS (underlined: linker peptide sequence of anti-TNF DVD-Ig).
    Figure Legend Snippet: TmTNF-dependent vesicular trafficking of humanized anti-TNFs and detection of cell surface anti-TNF peptides in dendritic cells. (A) Compartmentalization of anti-TNF: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and the cells ‘pulsed’ on ice using Alexa 488-conjugated anti-TNF (0 min, ice). The fluorescent label on the anti-TNF was ‘chased’ for up to 1 h at 37°C. Endosomes and lysosomes were stained with Alexa 647-conjuated anti-EEA1 and anti-LAMP1, respectively. Nucleus was stained with DAPI. Images were acquired using confocal microscopy. (B) Identification of cell surface-associated anti-TNF peptides: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and incubated with either an anti-TNF mAb or a DVD-Ig containing one anti-TNF domain for 6 h. Cell surface displayed peptides were eluted under mild acidic conditions and the identity of anti-TNF peptides was obtained using LC-MS/MS. (C) Identification of HLA-DR-associated anti-TNF peptides: Day 5 human CD14+ monocyte-derived DCs were treated with LPS for 2 h and incubated with anti-TNF for up to 8 h. HLA-DR was immunoprecipitated and the identity of HLA-DR-associated anti-TNF peptides was obtained using LC-MS/MS (underlined: linker peptide sequence of anti-TNF DVD-Ig).

    Techniques Used: Derivative Assay, Staining, Confocal Microscopy, Incubation, Liquid Chromatography with Mass Spectroscopy, Immunoprecipitation, Sequencing

    anti eea1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eea1
    Indirect immunofluorescence confocal microscopy with anti-PC1/3 (green), (A) anti-TGN46 (red), (B) <t>anti-EEA1</t> (red) and (C) anti-LAMP1 (red). A. The arrow indicates the TGN region where PC1/3 and TGN46 co-localize. C. The left arrow indicates the phagocytic structure and the lower right arrow indicates the lysosome-like structure where PC1/3 and LAMP1 are partially co-localized.
    Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eea1/product/Cell Signaling Technology Inc
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    Images

    1) Product Images from "Proprotein Convertase 1/3 (PC1/3) in the Rat Alveolar Macrophage Cell Line NR8383: Localization, Trafficking and Effects on Cytokine Secretion"

    Article Title: Proprotein Convertase 1/3 (PC1/3) in the Rat Alveolar Macrophage Cell Line NR8383: Localization, Trafficking and Effects on Cytokine Secretion

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0061557

    Indirect immunofluorescence confocal microscopy with anti-PC1/3 (green), (A) anti-TGN46 (red), (B) anti-EEA1 (red) and (C) anti-LAMP1 (red). A. The arrow indicates the TGN region where PC1/3 and TGN46 co-localize. C. The left arrow indicates the phagocytic structure and the lower right arrow indicates the lysosome-like structure where PC1/3 and LAMP1 are partially co-localized.
    Figure Legend Snippet: Indirect immunofluorescence confocal microscopy with anti-PC1/3 (green), (A) anti-TGN46 (red), (B) anti-EEA1 (red) and (C) anti-LAMP1 (red). A. The arrow indicates the TGN region where PC1/3 and TGN46 co-localize. C. The left arrow indicates the phagocytic structure and the lower right arrow indicates the lysosome-like structure where PC1/3 and LAMP1 are partially co-localized.

    Techniques Used: Immunofluorescence, Confocal Microscopy

    Confocal images of NR8383 cells stably transformed with control shRNA (NT) and shRNA directed against PC1/3 labeled with (A) early endosome marker EEA1 or RAB5. (D) baso-lateral membrane transport marker RAB8 and (F) late endosome/lysosome marker RAB7. In (A), arrows indicate punctate labeling near the plasma membrane. Insets represent 3× magnification. (B–C, G) EEA1 (B), RAB5 (C) and RAB7 (G) labelled vesicles area was estimated using ImageJ software. Vesicles where considered as ovals and area was calculated using the following formula: π×a×b where a and b are the two largest diameters. E. The ratio between peripheral and total integrated intensity of RAB8 labeling is represented. H. RAB7 peri nuclear integrated intensity is represented. * = 0.05 ** = 0.001 ***<0.0001, p-values, Student’s t-test. Quantification was performed on randomly selected field of view on two independent shRNA cell lines n = 4–6.
    Figure Legend Snippet: Confocal images of NR8383 cells stably transformed with control shRNA (NT) and shRNA directed against PC1/3 labeled with (A) early endosome marker EEA1 or RAB5. (D) baso-lateral membrane transport marker RAB8 and (F) late endosome/lysosome marker RAB7. In (A), arrows indicate punctate labeling near the plasma membrane. Insets represent 3× magnification. (B–C, G) EEA1 (B), RAB5 (C) and RAB7 (G) labelled vesicles area was estimated using ImageJ software. Vesicles where considered as ovals and area was calculated using the following formula: π×a×b where a and b are the two largest diameters. E. The ratio between peripheral and total integrated intensity of RAB8 labeling is represented. H. RAB7 peri nuclear integrated intensity is represented. * = 0.05 ** = 0.001 ***<0.0001, p-values, Student’s t-test. Quantification was performed on randomly selected field of view on two independent shRNA cell lines n = 4–6.

    Techniques Used: Stable Transfection, Transformation Assay, shRNA, Labeling, Marker, Software

    Representative western blots (20 µg of protein) and gels optic density (OD) quantification showing the relative levels of vesicle trafficking markers EEA1 (A), RAB5 (B), RAB8 (C), RAB7 (D), RAB9 (E) and RAB11 (F) between NR8383 cells expressing control shRNA (NT) and those expressing shRNA directed against PC1/3 (SH). The actin loading control is included. * = 0.05 ** = 0.001 ***<0.0001 p-values, Student’s t-test n = 5–8.
    Figure Legend Snippet: Representative western blots (20 µg of protein) and gels optic density (OD) quantification showing the relative levels of vesicle trafficking markers EEA1 (A), RAB5 (B), RAB8 (C), RAB7 (D), RAB9 (E) and RAB11 (F) between NR8383 cells expressing control shRNA (NT) and those expressing shRNA directed against PC1/3 (SH). The actin loading control is included. * = 0.05 ** = 0.001 ***<0.0001 p-values, Student’s t-test n = 5–8.

    Techniques Used: Western Blot, Expressing, shRNA

    anti eea1 rabbit monoclonal antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti eea1 rabbit monoclonal antibody
    (A) Two different concentrations of the FITC-labeled heptameric Z EGFR targeting ligands were incubated with A431 cells for 2 h at 37°C. (B) FITC labeled heptameric Z HER2 targeting ligands at two concentrations were incubated with SK-OV3 cells for 2 h at 37°C. <t>EEA1</t> proteins were detected by Alexa 555-conjugated secondary antibody. Top left panels: cell nuclei stained with DAPI (blue); Top right panels: FITC labeled heptamer (green); bottom left panels: EEA1 antibody (red); bottom right panels: merged image of the three stainings.
    Anti Eea1 Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eea1 rabbit monoclonal antibody/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti eea1 rabbit monoclonal antibody - by Bioz Stars, 2023-02
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    Images

    1) Product Images from "Heptameric Targeting Ligands against EGFR and HER2 with High Stability and Avidity"

    Article Title: Heptameric Targeting Ligands against EGFR and HER2 with High Stability and Avidity

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0043077

    (A) Two different concentrations of the FITC-labeled heptameric Z EGFR targeting ligands were incubated with A431 cells for 2 h at 37°C. (B) FITC labeled heptameric Z HER2 targeting ligands at two concentrations were incubated with SK-OV3 cells for 2 h at 37°C. EEA1 proteins were detected by Alexa 555-conjugated secondary antibody. Top left panels: cell nuclei stained with DAPI (blue); Top right panels: FITC labeled heptamer (green); bottom left panels: EEA1 antibody (red); bottom right panels: merged image of the three stainings.
    Figure Legend Snippet: (A) Two different concentrations of the FITC-labeled heptameric Z EGFR targeting ligands were incubated with A431 cells for 2 h at 37°C. (B) FITC labeled heptameric Z HER2 targeting ligands at two concentrations were incubated with SK-OV3 cells for 2 h at 37°C. EEA1 proteins were detected by Alexa 555-conjugated secondary antibody. Top left panels: cell nuclei stained with DAPI (blue); Top right panels: FITC labeled heptamer (green); bottom left panels: EEA1 antibody (red); bottom right panels: merged image of the three stainings.

    Techniques Used: Labeling, Incubation, Staining

    anti eea1  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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    Structured Review

    Cell Signaling Technology Inc anti eea1
    Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eea1/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti eea1 - by Bioz Stars, 2023-02
    97/100 stars

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    Cell Signaling Technology Inc anti eea1
    Anti Eea1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc eea1
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    Cell Signaling Technology Inc rabbit monoclonal early endosome antigen 1
    Effects of GM1 treatment on PDGFR activation. (a) Western blot analysis and quantification of PDGFR- β activation. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1, as compared to hTSCs differentiated in GM1-free osteogenic medium (O.D.). Total proteins were extracted and analyzed with anti-phosphorylated-PDGFR- β (Tyr 751) antibody (green) and anti-PDGFR- β (28E1) antibody (red). <t>EEA1</t> expression was used as internal control. Data are means ± SD of four different experiments. (b, c) Gene expression analysis of the osteogenic markers ALP and osteocalcin by real-time PCR. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1 or 10 ng/ml PDGF-BB or with both 100 μ M GM1 and 10 ng/ml PDGF-BB. The results were compared to hTSCs differentiated in free osteogenic medium (O.D.). Ribosomal protein S14 gene was used as housekeeper. All data are means ± SD of three different experiments. The statistical analysis was determined by Student's t-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.
    Rabbit Monoclonal Early Endosome Antigen 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti eea1 rabbit monoclonal antibody
    (A) Two different concentrations of the FITC-labeled heptameric Z EGFR targeting ligands were incubated with A431 cells for 2 h at 37°C. (B) FITC labeled heptameric Z HER2 targeting ligands at two concentrations were incubated with SK-OV3 cells for 2 h at 37°C. <t>EEA1</t> proteins were detected by Alexa 555-conjugated secondary antibody. Top left panels: cell nuclei stained with DAPI (blue); Top right panels: FITC labeled heptamer (green); bottom left panels: EEA1 antibody (red); bottom right panels: merged image of the three stainings.
    Anti Eea1 Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti eea1 rabbit monoclonal antibody/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti eea1 rabbit monoclonal antibody - by Bioz Stars, 2023-02
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    Image Search Results


    Effects of GM1 treatment on PDGFR activation. (a) Western blot analysis and quantification of PDGFR- β activation. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1, as compared to hTSCs differentiated in GM1-free osteogenic medium (O.D.). Total proteins were extracted and analyzed with anti-phosphorylated-PDGFR- β (Tyr 751) antibody (green) and anti-PDGFR- β (28E1) antibody (red). EEA1 expression was used as internal control. Data are means ± SD of four different experiments. (b, c) Gene expression analysis of the osteogenic markers ALP and osteocalcin by real-time PCR. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1 or 10 ng/ml PDGF-BB or with both 100 μ M GM1 and 10 ng/ml PDGF-BB. The results were compared to hTSCs differentiated in free osteogenic medium (O.D.). Ribosomal protein S14 gene was used as housekeeper. All data are means ± SD of three different experiments. The statistical analysis was determined by Student's t-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Journal: Stem Cells International

    Article Title: GM1 Ganglioside Promotes Osteogenic Differentiation of Human Tendon Stem Cells

    doi: 10.1155/2018/4706943

    Figure Lengend Snippet: Effects of GM1 treatment on PDGFR activation. (a) Western blot analysis and quantification of PDGFR- β activation. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1, as compared to hTSCs differentiated in GM1-free osteogenic medium (O.D.). Total proteins were extracted and analyzed with anti-phosphorylated-PDGFR- β (Tyr 751) antibody (green) and anti-PDGFR- β (28E1) antibody (red). EEA1 expression was used as internal control. Data are means ± SD of four different experiments. (b, c) Gene expression analysis of the osteogenic markers ALP and osteocalcin by real-time PCR. hTSCs were differentiated toward osteoblasts in osteogenic medium supplemented with 100 μ M GM1 or 10 ng/ml PDGF-BB or with both 100 μ M GM1 and 10 ng/ml PDGF-BB. The results were compared to hTSCs differentiated in free osteogenic medium (O.D.). Ribosomal protein S14 gene was used as housekeeper. All data are means ± SD of three different experiments. The statistical analysis was determined by Student's t-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Article Snippet: After blocking the membranes with 5% ( w / v ) of nonfat dry milk in Tris-buffered saline-Tween 0.1% (TBS-T) for 1 hour at room temperature, they were incubated overnight at 4°C with the following primary antibodies: rabbit phospho-PDGFR- β , 1 : 1000 dilution (Y751, Cell Signaling); rabbit PDGFR- β , 1 : 1000 dilution (Cell Signaling); and rabbit monoclonal early endosome antigen 1 (EEA1), 1 : 1000 dilution (Cell Signaling).

    Techniques: Activation Assay, Western Blot, Expressing, Real-time Polymerase Chain Reaction

    (A) Two different concentrations of the FITC-labeled heptameric Z EGFR targeting ligands were incubated with A431 cells for 2 h at 37°C. (B) FITC labeled heptameric Z HER2 targeting ligands at two concentrations were incubated with SK-OV3 cells for 2 h at 37°C. EEA1 proteins were detected by Alexa 555-conjugated secondary antibody. Top left panels: cell nuclei stained with DAPI (blue); Top right panels: FITC labeled heptamer (green); bottom left panels: EEA1 antibody (red); bottom right panels: merged image of the three stainings.

    Journal: PLoS ONE

    Article Title: Heptameric Targeting Ligands against EGFR and HER2 with High Stability and Avidity

    doi: 10.1371/journal.pone.0043077

    Figure Lengend Snippet: (A) Two different concentrations of the FITC-labeled heptameric Z EGFR targeting ligands were incubated with A431 cells for 2 h at 37°C. (B) FITC labeled heptameric Z HER2 targeting ligands at two concentrations were incubated with SK-OV3 cells for 2 h at 37°C. EEA1 proteins were detected by Alexa 555-conjugated secondary antibody. Top left panels: cell nuclei stained with DAPI (blue); Top right panels: FITC labeled heptamer (green); bottom left panels: EEA1 antibody (red); bottom right panels: merged image of the three stainings.

    Article Snippet: The cells were then incubated with the anti-EEA1 rabbit monoclonal antibody (1:200) (Cell Signaling Technology, Danvers, MA) overnight at 4°C.

    Techniques: Labeling, Incubation, Staining