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  • 5a1  (Prospec)
    79
    Prospec 5a1
    5a1, supplied by Prospec, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5a1/product/Prospec
    Average 79 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    5a1 - by Bioz Stars, 2019-10
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    5a1  (ATCC)
    77
    ATCC 5a1
    5a1, supplied by ATCC, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5a1/product/ATCC
    Average 77 stars, based on 1 article reviews
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    5a1 - by Bioz Stars, 2019-10
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    77
    Novartis 5a1
    Toxicity of <t>5A1</t> on MC can be rescued A. Metabolic activity of MC is increased in co-culture with breast cancer cells or IL-34 treatment, analyzed by MTT reduction (mean ± SD, n = 4, *** P
    5a1, supplied by Novartis, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5a1/product/Novartis
    Average 77 stars, based on 1 article reviews
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    5a1 - by Bioz Stars, 2019-10
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    86
    Abnova aaas monoclonal antibody m02 clone 5a1
    Toxicity of <t>5A1</t> on MC can be rescued A. Metabolic activity of MC is increased in co-culture with breast cancer cells or IL-34 treatment, analyzed by MTT reduction (mean ± SD, n = 4, *** P
    Aaas Monoclonal Antibody M02 Clone 5a1, supplied by Abnova, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aaas monoclonal antibody m02 clone 5a1/product/Abnova
    Average 86 stars, based on 9 article reviews
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    aaas monoclonal antibody m02 clone 5a1 - by Bioz Stars, 2019-10
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    82
    Becton Dickinson mouse anti eif 5a1
    Dohh -mediated hypusination of <t>eIF-5A1</t> is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.
    Mouse Anti Eif 5a1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 82/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti eif 5a1/product/Becton Dickinson
    Average 82 stars, based on 6 article reviews
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    81
    Bio X Cell αcsf1 5a1 antibodies
    Dohh -mediated hypusination of <t>eIF-5A1</t> is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.
    αcsf1 5a1 Antibodies, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 81/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 81 stars, based on 5 article reviews
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    79
    Bio X Cell anti csf1 5a1 monoclonal antibody
    Dohh -mediated hypusination of <t>eIF-5A1</t> is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.
    Anti Csf1 5a1 Monoclonal Antibody, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Cell Signaling Technology Inc anti cleaved caspase3 asp175 5a1 rabbit mab
    Dohh -mediated hypusination of <t>eIF-5A1</t> is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.
    Anti Cleaved Caspase3 Asp175 5a1 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 79/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cleaved caspase3 asp175 5a1 rabbit mab/product/Cell Signaling Technology Inc
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    76
    Cell Signaling Technology Inc cleaved caspase 3 5a1
    Dohh -mediated hypusination of <t>eIF-5A1</t> is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.
    Cleaved Caspase 3 5a1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cleaved caspase 3 5a1/product/Cell Signaling Technology Inc
    Average 76 stars, based on 1 article reviews
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    76
    Novus Biologicals anti eif 5a1
    Dohh -mediated hypusination of <t>eIF-5A1</t> is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.
    Anti Eif 5a1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    Novus Biologicals rabbit anti eif 5a1
    Establishment of a protein fragment complementation assay (PCA) assay to validate newly identified PPI. A , Mechanism of the split-YFP PCA assay: upon interaction of proteins A and C, the YFP fragments can fold into YFP's native state, leading to fluorescence activity ( B ) Flow cytometric evaluation of previously known PPI and negative controls involving <t>eIF-5A1-</t> and MCFD2-YFP 1 . Corresponding YFP 2 -coupled protein is indicated by shape. MCFD2-YFP 1 and ERGIC-53-YFP 2 are known to interact ( 31 ) and show MCFD2-YFP 1 is expressed and functional, confirming its applicability as negative control. Values represent mean fluorescence of viable cotransfected cells. C , Representative microscopic pictures of a control combination (eIF-5A2 + MCFD2) and a known interaction (eIF-5A2 + DOHH). The elevated background of the control originates from prolonged exposure. D–G , Flow cytometric quantification of PCA experiments with combinations of known and newly identified putative interaction partners. Bars represent mean fluorescence of two independent runs of duplicates ( n = 4) normalized to negative control experiments (gray dashed lines). Error bars represent standard error of the mean (S.E.). Asterisks (*) represent statistically significant variation from the respective negative control (* p
    Rabbit Anti Eif 5a1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Toxicity of 5A1 on MC can be rescued A. Metabolic activity of MC is increased in co-culture with breast cancer cells or IL-34 treatment, analyzed by MTT reduction (mean ± SD, n = 4, *** P

    Journal: Oncotarget

    Article Title: Anti-CSF-1 treatment is effective to prevent carcinoma invasion induced by monocyte-derived cells but scarcely by microglia

    doi:

    Figure Lengend Snippet: Toxicity of 5A1 on MC can be rescued A. Metabolic activity of MC is increased in co-culture with breast cancer cells or IL-34 treatment, analyzed by MTT reduction (mean ± SD, n = 4, *** P

    Article Snippet: The anti-CSF1 antibody, 5A1, was provided by Novartis.

    Techniques: Activity Assay, Co-Culture Assay, MTT Assay

    Differing cytotoxicity of 5A1 on distinct macrophage populations is correlated with differing growth factor expression MC A. and MG B. were treated with 0 μg/ml (circle), 2.5 μg/ml (square), 10 μg/ml (triangle) and 50 μg/ml (inverse triangle) 5A1. Cell proliferation was measured over 48 h using the xCELLigence system and is indicated as cell index. Shown is one representative result ( n = 3). C, D. qRT-PCR for CSF-1, IL-34 and its receptor CSF-1R in (C) MG (circle) and MC (square) and (D) the respective tissue of origin of these two macrophage populations, i.e. brain (circle) and bone marrow (BM, square) (GOI = gene of interest, HK = housekeeper, n.e. = not expressed, * P

    Journal: Oncotarget

    Article Title: Anti-CSF-1 treatment is effective to prevent carcinoma invasion induced by monocyte-derived cells but scarcely by microglia

    doi:

    Figure Lengend Snippet: Differing cytotoxicity of 5A1 on distinct macrophage populations is correlated with differing growth factor expression MC A. and MG B. were treated with 0 μg/ml (circle), 2.5 μg/ml (square), 10 μg/ml (triangle) and 50 μg/ml (inverse triangle) 5A1. Cell proliferation was measured over 48 h using the xCELLigence system and is indicated as cell index. Shown is one representative result ( n = 3). C, D. qRT-PCR for CSF-1, IL-34 and its receptor CSF-1R in (C) MG (circle) and MC (square) and (D) the respective tissue of origin of these two macrophage populations, i.e. brain (circle) and bone marrow (BM, square) (GOI = gene of interest, HK = housekeeper, n.e. = not expressed, * P

    Article Snippet: The anti-CSF1 antibody, 5A1, was provided by Novartis.

    Techniques: Expressing, Quantitative RT-PCR

    CSF-1R downstream-signaling is affected by 5A1 in MC but not in MG MC (left panel) and MG (right panel) were treated with 2.5 μg/ml 5A1, respectively. Expression of AKT, pAKT, S6 and pS6 was analyzed by western blot after 30 min of treatment. All western blots were repeated at least three times, shown is one representative example.

    Journal: Oncotarget

    Article Title: Anti-CSF-1 treatment is effective to prevent carcinoma invasion induced by monocyte-derived cells but scarcely by microglia

    doi:

    Figure Lengend Snippet: CSF-1R downstream-signaling is affected by 5A1 in MC but not in MG MC (left panel) and MG (right panel) were treated with 2.5 μg/ml 5A1, respectively. Expression of AKT, pAKT, S6 and pS6 was analyzed by western blot after 30 min of treatment. All western blots were repeated at least three times, shown is one representative example.

    Article Snippet: The anti-CSF1 antibody, 5A1, was provided by Novartis.

    Techniques: Expressing, Western Blot

    5A1 decreases MG- and MC-induced invasiveness of MCF-7 and 410.4 cells Microinvasion assay of tumor cells alone (white bars) and in co-culture with MC A, B. or MG C, D. in the absence (gray bars) or presence (black bars) of 2.5 μg/ml 5A1. MC- and MG-induced invasiveness of both MCF-7 (A), (C) and 410.4 (B), (D) is significantly decreased by 5A1. However MG-induced invasion is blocked by 5A1 to a lower extent. Invasiveness is indicated as the percentage of the control tumor cells alone (mean ± SD, n ≥ 4, * P

    Journal: Oncotarget

    Article Title: Anti-CSF-1 treatment is effective to prevent carcinoma invasion induced by monocyte-derived cells but scarcely by microglia

    doi:

    Figure Lengend Snippet: 5A1 decreases MG- and MC-induced invasiveness of MCF-7 and 410.4 cells Microinvasion assay of tumor cells alone (white bars) and in co-culture with MC A, B. or MG C, D. in the absence (gray bars) or presence (black bars) of 2.5 μg/ml 5A1. MC- and MG-induced invasiveness of both MCF-7 (A), (C) and 410.4 (B), (D) is significantly decreased by 5A1. However MG-induced invasion is blocked by 5A1 to a lower extent. Invasiveness is indicated as the percentage of the control tumor cells alone (mean ± SD, n ≥ 4, * P

    Article Snippet: The anti-CSF1 antibody, 5A1, was provided by Novartis.

    Techniques: Co-Culture Assay

    Anti-CSF-1 antibody 5A1 does not exert cytotoxic effects on tested breast cancer cells A, B. Metabolic activity of MCF-7 (A) and MDA-MB231 (B) was analyzed 96 h after treatment with 5A1 by measuring MTT reduction (mean ± SD, n ≥ 6). C, D. MCF-7 (C) and MDA-MB231 (D) were treated with 0 μg/ml (circle), 2.5 μg/ml (square), 10 μg/ml (triangle) and 50 μg/ml (inverse triangle) 5A1. Cell proliferation was measured over 48 h using the xCELLigence system and is indicated as cell index. E, F. ECM-based migration assays for MCF-7 and MDA-MB231 over 48 h in the absence (gray bars, left pictures) and presence (black bars, right pictures) of 25 μg/ml 5A1 (mean ± SD, n = 4). Scale bars indicate 200 μm.

    Journal: Oncotarget

    Article Title: Anti-CSF-1 treatment is effective to prevent carcinoma invasion induced by monocyte-derived cells but scarcely by microglia

    doi:

    Figure Lengend Snippet: Anti-CSF-1 antibody 5A1 does not exert cytotoxic effects on tested breast cancer cells A, B. Metabolic activity of MCF-7 (A) and MDA-MB231 (B) was analyzed 96 h after treatment with 5A1 by measuring MTT reduction (mean ± SD, n ≥ 6). C, D. MCF-7 (C) and MDA-MB231 (D) were treated with 0 μg/ml (circle), 2.5 μg/ml (square), 10 μg/ml (triangle) and 50 μg/ml (inverse triangle) 5A1. Cell proliferation was measured over 48 h using the xCELLigence system and is indicated as cell index. E, F. ECM-based migration assays for MCF-7 and MDA-MB231 over 48 h in the absence (gray bars, left pictures) and presence (black bars, right pictures) of 25 μg/ml 5A1 (mean ± SD, n = 4). Scale bars indicate 200 μm.

    Article Snippet: The anti-CSF1 antibody, 5A1, was provided by Novartis.

    Techniques: Activity Assay, Multiple Displacement Amplification, MTT Assay, Migration

    Dohh -mediated hypusination of eIF-5A1 is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.

    Journal:

    Article Title:

    doi: 10.1074/jbc.M115.664490

    Figure Lengend Snippet: Dohh -mediated hypusination of eIF-5A1 is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.

    Article Snippet: After washing, residual paraformaldehyde was quenched with 50 m m NH4 Cl in PBS for 10 min. For permeabilization, the cells were treated for 5 min with 0.1% (v/v) Triton X-100 in PBS, and then the samples were blocked with 0.5% (w/v) BSA in PBS for 30 min. Immunostaining was performed with mouse anti-eIF-5A1 (clone 26/eIF-5a; BD Biosciences) and Alexa Fluor 488-labeled goat anti-mouse antibody (Life Technologies, Inc.).

    Techniques: Mouse Assay

    Knockdown of the hypusine modification enzymes affect localization and post-translational modification of eIF-5A1. A, immunofluorescence for endogenous eIF-5A1 in a Dhs or Dohh -deficient cellular background, respectively. Upper panels depict eIF-5A1 fluorescence

    Journal:

    Article Title:

    doi: 10.1074/jbc.M115.664490

    Figure Lengend Snippet: Knockdown of the hypusine modification enzymes affect localization and post-translational modification of eIF-5A1. A, immunofluorescence for endogenous eIF-5A1 in a Dhs or Dohh -deficient cellular background, respectively. Upper panels depict eIF-5A1 fluorescence

    Article Snippet: After washing, residual paraformaldehyde was quenched with 50 m m NH4 Cl in PBS for 10 min. For permeabilization, the cells were treated for 5 min with 0.1% (v/v) Triton X-100 in PBS, and then the samples were blocked with 0.5% (w/v) BSA in PBS for 30 min. Immunostaining was performed with mouse anti-eIF-5A1 (clone 26/eIF-5a; BD Biosciences) and Alexa Fluor 488-labeled goat anti-mouse antibody (Life Technologies, Inc.).

    Techniques: Modification, Immunofluorescence, Fluorescence

    Loss of Dohh reveals histological anomalies. A, two-dimensional Western blot for eIF-5A1 in spleen tissue after 4-OHT treatment in vivo . Spleen was isolated from mice showing more than 20% weight loss. Colored arrows in the representative blots correspond

    Journal:

    Article Title:

    doi: 10.1074/jbc.M115.664490

    Figure Lengend Snippet: Loss of Dohh reveals histological anomalies. A, two-dimensional Western blot for eIF-5A1 in spleen tissue after 4-OHT treatment in vivo . Spleen was isolated from mice showing more than 20% weight loss. Colored arrows in the representative blots correspond

    Article Snippet: After washing, residual paraformaldehyde was quenched with 50 m m NH4 Cl in PBS for 10 min. For permeabilization, the cells were treated for 5 min with 0.1% (v/v) Triton X-100 in PBS, and then the samples were blocked with 0.5% (w/v) BSA in PBS for 30 min. Immunostaining was performed with mouse anti-eIF-5A1 (clone 26/eIF-5a; BD Biosciences) and Alexa Fluor 488-labeled goat anti-mouse antibody (Life Technologies, Inc.).

    Techniques: Western Blot, In Vivo, Isolation, Mouse Assay

    Dohh -mediated hypusination of eIF-5A1 is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.

    Journal:

    Article Title:

    doi: 10.1074/jbc.M115.664490

    Figure Lengend Snippet: Dohh -mediated hypusination of eIF-5A1 is essential in adult mice. A–D, analysis of adult Dohh +/+ , Dohh p/+ , and Dohh p/p -CAG-cre/Esr1 + -positive mice after 4-OHT administration resulting from a breeding according to the scheme in , B and C.

    Article Snippet: One- and two-dimensional Western blot analyses were carried out as described before ( , ) using anti-eIF-5A1 (Novus Biologicals, Littleton, CO) and anti-eIF-5A2 (Abcam, Cambridge, UK).

    Techniques: Mouse Assay

    Knockdown of the hypusine modification enzymes affect localization and post-translational modification of eIF-5A1. A, immunofluorescence for endogenous eIF-5A1 in a Dhs or Dohh -deficient cellular background, respectively. Upper panels depict eIF-5A1 fluorescence

    Journal:

    Article Title:

    doi: 10.1074/jbc.M115.664490

    Figure Lengend Snippet: Knockdown of the hypusine modification enzymes affect localization and post-translational modification of eIF-5A1. A, immunofluorescence for endogenous eIF-5A1 in a Dhs or Dohh -deficient cellular background, respectively. Upper panels depict eIF-5A1 fluorescence

    Article Snippet: One- and two-dimensional Western blot analyses were carried out as described before ( , ) using anti-eIF-5A1 (Novus Biologicals, Littleton, CO) and anti-eIF-5A2 (Abcam, Cambridge, UK).

    Techniques: Modification, Immunofluorescence, Fluorescence

    Loss of Dohh reveals histological anomalies. A, two-dimensional Western blot for eIF-5A1 in spleen tissue after 4-OHT treatment in vivo . Spleen was isolated from mice showing more than 20% weight loss. Colored arrows in the representative blots correspond

    Journal:

    Article Title:

    doi: 10.1074/jbc.M115.664490

    Figure Lengend Snippet: Loss of Dohh reveals histological anomalies. A, two-dimensional Western blot for eIF-5A1 in spleen tissue after 4-OHT treatment in vivo . Spleen was isolated from mice showing more than 20% weight loss. Colored arrows in the representative blots correspond

    Article Snippet: One- and two-dimensional Western blot analyses were carried out as described before ( , ) using anti-eIF-5A1 (Novus Biologicals, Littleton, CO) and anti-eIF-5A2 (Abcam, Cambridge, UK).

    Techniques: Western Blot, In Vivo, Isolation, Mouse Assay

    Establishment of a protein fragment complementation assay (PCA) assay to validate newly identified PPI. A , Mechanism of the split-YFP PCA assay: upon interaction of proteins A and C, the YFP fragments can fold into YFP's native state, leading to fluorescence activity ( B ) Flow cytometric evaluation of previously known PPI and negative controls involving eIF-5A1- and MCFD2-YFP 1 . Corresponding YFP 2 -coupled protein is indicated by shape. MCFD2-YFP 1 and ERGIC-53-YFP 2 are known to interact ( 31 ) and show MCFD2-YFP 1 is expressed and functional, confirming its applicability as negative control. Values represent mean fluorescence of viable cotransfected cells. C , Representative microscopic pictures of a control combination (eIF-5A2 + MCFD2) and a known interaction (eIF-5A2 + DOHH). The elevated background of the control originates from prolonged exposure. D–G , Flow cytometric quantification of PCA experiments with combinations of known and newly identified putative interaction partners. Bars represent mean fluorescence of two independent runs of duplicates ( n = 4) normalized to negative control experiments (gray dashed lines). Error bars represent standard error of the mean (S.E.). Asterisks (*) represent statistically significant variation from the respective negative control (* p

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title:

    doi: 10.1074/mcp.M112.019059

    Figure Lengend Snippet: Establishment of a protein fragment complementation assay (PCA) assay to validate newly identified PPI. A , Mechanism of the split-YFP PCA assay: upon interaction of proteins A and C, the YFP fragments can fold into YFP's native state, leading to fluorescence activity ( B ) Flow cytometric evaluation of previously known PPI and negative controls involving eIF-5A1- and MCFD2-YFP 1 . Corresponding YFP 2 -coupled protein is indicated by shape. MCFD2-YFP 1 and ERGIC-53-YFP 2 are known to interact ( 31 ) and show MCFD2-YFP 1 is expressed and functional, confirming its applicability as negative control. Values represent mean fluorescence of viable cotransfected cells. C , Representative microscopic pictures of a control combination (eIF-5A2 + MCFD2) and a known interaction (eIF-5A2 + DOHH). The elevated background of the control originates from prolonged exposure. D–G , Flow cytometric quantification of PCA experiments with combinations of known and newly identified putative interaction partners. Bars represent mean fluorescence of two independent runs of duplicates ( n = 4) normalized to negative control experiments (gray dashed lines). Error bars represent standard error of the mean (S.E.). Asterisks (*) represent statistically significant variation from the respective negative control (* p

    Article Snippet: The employed antibodies were rabbit anti-eIF-5A1 (Novus Biologicals, Littleton, CA), rabbit anti-Myc-tag (Cell Signaling, Boston, MA), and anti-rabbit-HRP (Cell Signaling).

    Techniques: Protein-Fragment Complementation Assay, Fluorescence, Activity Assay, Flow Cytometry, Functional Assay, Negative Control

    Specificity and subcellular localization of observed eIF-5A1/DHS-complexes. Fluorescence microscopy images showing representative YFP-positive cells from PCA experiments as indicated on the panels. Green channel, reconstituted YFP; red channel, mRFP or Alexa633 (immunolabeling experiments); blue channel, Cer2. A , Wild type eIF-5A1, the eIF-5A1 K50R mutant and the N-terminal domain of eIF-5A1 show YFP speckles upon binding of DHS in PCA experiments. The eIF-5A1 G52A mutant, the C-terminal domain of eIF-5A1 and addition of 20 μ m GC7 disrupt this localization of eIF-5A1/DHS PCA complexes. B , A fusion protein from fluorescent proteins mRFP or Cer2, respectively, and TSG101 (member of the ESCRT-I protein family and marker for MVBs) showed significant colocalization with eIF-5A1/DHS PCA complexes. Most double-positive cells showed more speckles of eIF-5A1/DHS PCA complex localization than MVBs, but the majority of labeled MVBs colocalized to eIF-5A1/DHS PCA complexes. C , Immunolabeled EEA1 (marker for early endosomes) also showed significant colocalization with eIF-5A1/DHS PCA complexes.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title:

    doi: 10.1074/mcp.M112.019059

    Figure Lengend Snippet: Specificity and subcellular localization of observed eIF-5A1/DHS-complexes. Fluorescence microscopy images showing representative YFP-positive cells from PCA experiments as indicated on the panels. Green channel, reconstituted YFP; red channel, mRFP or Alexa633 (immunolabeling experiments); blue channel, Cer2. A , Wild type eIF-5A1, the eIF-5A1 K50R mutant and the N-terminal domain of eIF-5A1 show YFP speckles upon binding of DHS in PCA experiments. The eIF-5A1 G52A mutant, the C-terminal domain of eIF-5A1 and addition of 20 μ m GC7 disrupt this localization of eIF-5A1/DHS PCA complexes. B , A fusion protein from fluorescent proteins mRFP or Cer2, respectively, and TSG101 (member of the ESCRT-I protein family and marker for MVBs) showed significant colocalization with eIF-5A1/DHS PCA complexes. Most double-positive cells showed more speckles of eIF-5A1/DHS PCA complex localization than MVBs, but the majority of labeled MVBs colocalized to eIF-5A1/DHS PCA complexes. C , Immunolabeled EEA1 (marker for early endosomes) also showed significant colocalization with eIF-5A1/DHS PCA complexes.

    Article Snippet: The employed antibodies were rabbit anti-eIF-5A1 (Novus Biologicals, Littleton, CA), rabbit anti-Myc-tag (Cell Signaling, Boston, MA), and anti-rabbit-HRP (Cell Signaling).

    Techniques: Fluorescence, Microscopy, Immunolabeling, Mutagenesis, Binding Assay, Marker, Labeling