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Proteintech anti e1b ap5
The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, <t>E1B-AP5,</t> and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .
Anti E1b Ap5, supplied by Proteintech, used in various techniques. Bioz Stars score: 85/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti e1b ap5 - by Bioz Stars, 2020-09
85/100 stars

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1) Product Images from "Influenza virus targets the mRNA export machinery and the nuclear pore complex"

Article Title: Influenza virus targets the mRNA export machinery and the nuclear pore complex

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0610977104

The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .
Figure Legend Snippet: The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .

Techniques Used: Incubation, Recombinant, SDS Page, Expressing, Infection, Labeling

2) Product Images from "Vesicular stomatitis virus inhibits mitotic progression and triggers cell death"

Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death

Journal: EMBO Reports

doi: 10.1038/embor.2009.179

VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, EIB-AP5 or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using E1B-AP5 antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.
Figure Legend Snippet: VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, EIB-AP5 or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using E1B-AP5 antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.

Techniques Used: Incubation, Recombinant, SDS Page, Immunoprecipitation, Immunofluorescence, Microscopy, Polyacrylamide Gel Electrophoresis

3) Product Images from "Influenza virus targets the mRNA export machinery and the nuclear pore complex"

Article Title: Influenza virus targets the mRNA export machinery and the nuclear pore complex

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0610977104

The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .
Figure Legend Snippet: The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .

Techniques Used: Incubation, Recombinant, SDS Page, Expressing, Infection, Labeling

Selective impairment of mRNA nuclear export in Rae1 and Nup98 mutant cells. Nuclear (N) and cytoplasmic (C) RNA was isolated from Rae1 +/+ Nup98 +/+ , Rae1 +/+ Nup98 +/− , Rae1 +/− Nup98 +/+ , and Rae1 +/− Nup98 +/− MEFs. Relative mRNA levels were quantified by real-time RT-PCR using gene-specific primers and normalized to a set of five housekeeping genes.
Figure Legend Snippet: Selective impairment of mRNA nuclear export in Rae1 and Nup98 mutant cells. Nuclear (N) and cytoplasmic (C) RNA was isolated from Rae1 +/+ Nup98 +/+ , Rae1 +/+ Nup98 +/− , Rae1 +/− Nup98 +/+ , and Rae1 +/− Nup98 +/− MEFs. Relative mRNA levels were quantified by real-time RT-PCR using gene-specific primers and normalized to a set of five housekeeping genes.

Techniques Used: Mutagenesis, Isolation, Quantitative RT-PCR

Low levels of Rae1 and Nup98 induce higher susceptibility to influenza virus-mediated cell death and an increase in viral replication. ( a and b ) Rae1 +/+ Nup98 +/+ , Rae1 +/− Nup98 +/+ , Rae1 +/+ Nup98 +/− , and Rae1 +/− Nup98 +/− MEFs were infected with A/WS/33 influenza virus, and cell viability was determined by comparing and quantifying bright-field microscopy (gray), DAPI (blue), and exclusion of 2 mM ethidium homodimer-1 (red). ( c ) The number of influenza viral particles was measured in the supernatants of the cells in a by using the hemaglutinin assay.
Figure Legend Snippet: Low levels of Rae1 and Nup98 induce higher susceptibility to influenza virus-mediated cell death and an increase in viral replication. ( a and b ) Rae1 +/+ Nup98 +/+ , Rae1 +/− Nup98 +/+ , Rae1 +/+ Nup98 +/− , and Rae1 +/− Nup98 +/− MEFs were infected with A/WS/33 influenza virus, and cell viability was determined by comparing and quantifying bright-field microscopy (gray), DAPI (blue), and exclusion of 2 mM ethidium homodimer-1 (red). ( c ) The number of influenza viral particles was measured in the supernatants of the cells in a by using the hemaglutinin assay.

Techniques Used: Infection, Microscopy

The mRNA export inhibition induced by NS1 is reverted by increased levels of mRNA export factors. ( a and b ) Luciferase reporter gene expression assays were performed with 293T cells by cotransfection of reporter plasmids and plasmids encoding NXF1, p15, Rae1, Nup98, and Nup96, as indicated. ( c ) HeLa cells were transfected with a plasmid encoding myc-NS1 alone or cotransfected with plasmids encoding myc-NS1, GFP-NXF1, and GFP-p15. Cells were subjected to immunofluorescence with anti-myc antibody (red) followed by oligo(dT) in situ hybridization (blue). Green shows GFP-NXF1 and GFP-p15.
Figure Legend Snippet: The mRNA export inhibition induced by NS1 is reverted by increased levels of mRNA export factors. ( a and b ) Luciferase reporter gene expression assays were performed with 293T cells by cotransfection of reporter plasmids and plasmids encoding NXF1, p15, Rae1, Nup98, and Nup96, as indicated. ( c ) HeLa cells were transfected with a plasmid encoding myc-NS1 alone or cotransfected with plasmids encoding myc-NS1, GFP-NXF1, and GFP-p15. Cells were subjected to immunofluorescence with anti-myc antibody (red) followed by oligo(dT) in situ hybridization (blue). Green shows GFP-NXF1 and GFP-p15.

Techniques Used: Inhibition, Luciferase, Expressing, Cotransfection, Transfection, Plasmid Preparation, Immunofluorescence, In Situ Hybridization

4) Product Images from "Influenza virus targets the mRNA export machinery and the nuclear pore complex"

Article Title: Influenza virus targets the mRNA export machinery and the nuclear pore complex

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0610977104

The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .
Figure Legend Snippet: The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .

Techniques Used: Incubation, Recombinant, SDS Page, Expressing, Infection, Labeling

Influenza virus inhibits poly(A) RNA nuclear export. ( a ) MDCK cells were mock infected or infected with A/WS/33 influenza virus at an MOI of 1 for 6 and 24 h. Immunofluorescence using antibodies against influenza proteins (green) and oligo(dT) in situ hybridization (red) was performed. ( b ) Expression of influenza proteins in MDCKs. Cell extracts from MDCK cells infected with A/WS/33 at an MOI 1 for the indicated time points were subjected to immunoblot analysis with anti-influenza protein antibodies.
Figure Legend Snippet: Influenza virus inhibits poly(A) RNA nuclear export. ( a ) MDCK cells were mock infected or infected with A/WS/33 influenza virus at an MOI of 1 for 6 and 24 h. Immunofluorescence using antibodies against influenza proteins (green) and oligo(dT) in situ hybridization (red) was performed. ( b ) Expression of influenza proteins in MDCKs. Cell extracts from MDCK cells infected with A/WS/33 at an MOI 1 for the indicated time points were subjected to immunoblot analysis with anti-influenza protein antibodies.

Techniques Used: Infection, Immunofluorescence, In Situ Hybridization, Expressing

5) Product Images from "Stability of a Long Noncoding Viral RNA Depends on a 9-nt Core Element at the RNA 5' End to Interact with Viral ORF57 and Cellular PABPC1"

Article Title: Stability of a Long Noncoding Viral RNA Depends on a 9-nt Core Element at the RNA 5' End to Interact with Viral ORF57 and Cellular PABPC1

Journal: International Journal of Biological Sciences

doi:

MRE-II RNA interacts in vitro with ORF57 and other cellular proteins. ( A ) Mapping of an MRE binding site for ORF57. Shown on the top is a PAN MRE sequence lacking the MRE-I region in Fig. 4 A, with the 9-nt core of MRE-II loop sequence bolded and underlined and the nt positions of biotinylated RNA oligomers employed in RNA pull-down assays. A cell lysate prepared from TREx BCBL-1-Rta cells 39 induced with Dox for 24 h was used for the RNA pulldown assays with an indicated RNA oligomer. The RNA oligo oNP42 derived from vIL-6 RNA 27 which binds ORF57 was used as a positive control. ORF57 associated with RNA oligos in the pulldowns was immunoblotted using an anti-ORF57 antibody. The cell lysate (10%) before the pulldown was loaded as a Western blot control. ( B ) Biotinylated RNA affinity pulldown analysis by Western blot using anti-ORF57, PABPC1 and E1B-AP5 antibodies. Total cell extract from TREx BCBL-1-Rta (R) or -vector (V) cells induced by Dox for 24 h was used for the RNA pulldown assays with each biotinylated RNA oligomer. RNA oligos oNP41 and oNP42 derived from vIL-6 RNA 27 were used, respectively, as a negative and positive oligomer control for ORF57 interaction and 10% of the cell lysate from each cell line before the pulldown was loaded for Western blotting control. RNA oligo oJM68 is a copy of oJM35 with the point mutations in the MRE-II loop described in Fig. 4 B. Control beads indicate no RNA oligomer on the beads.
Figure Legend Snippet: MRE-II RNA interacts in vitro with ORF57 and other cellular proteins. ( A ) Mapping of an MRE binding site for ORF57. Shown on the top is a PAN MRE sequence lacking the MRE-I region in Fig. 4 A, with the 9-nt core of MRE-II loop sequence bolded and underlined and the nt positions of biotinylated RNA oligomers employed in RNA pull-down assays. A cell lysate prepared from TREx BCBL-1-Rta cells 39 induced with Dox for 24 h was used for the RNA pulldown assays with an indicated RNA oligomer. The RNA oligo oNP42 derived from vIL-6 RNA 27 which binds ORF57 was used as a positive control. ORF57 associated with RNA oligos in the pulldowns was immunoblotted using an anti-ORF57 antibody. The cell lysate (10%) before the pulldown was loaded as a Western blot control. ( B ) Biotinylated RNA affinity pulldown analysis by Western blot using anti-ORF57, PABPC1 and E1B-AP5 antibodies. Total cell extract from TREx BCBL-1-Rta (R) or -vector (V) cells induced by Dox for 24 h was used for the RNA pulldown assays with each biotinylated RNA oligomer. RNA oligos oNP41 and oNP42 derived from vIL-6 RNA 27 were used, respectively, as a negative and positive oligomer control for ORF57 interaction and 10% of the cell lysate from each cell line before the pulldown was loaded for Western blotting control. RNA oligo oJM68 is a copy of oJM35 with the point mutations in the MRE-II loop described in Fig. 4 B. Control beads indicate no RNA oligomer on the beads.

Techniques Used: In Vitro, Binding Assay, Sequencing, Derivative Assay, Positive Control, Western Blot, Plasmid Preparation

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    Proteintech e1b ap5
    VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, <t>EIB-AP5</t> or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using <t>E1B-AP5</t> antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.
    E1b Ap5, supplied by Proteintech, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e1b ap5/product/Proteintech
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    e1b ap5 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    85
    Proteintech anti e1b ap5
    The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, <t>E1B-AP5,</t> and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .
    Anti E1b Ap5, supplied by Proteintech, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti e1b ap5/product/Proteintech
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti e1b ap5 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    99
    Proteintech rabbit polyclonal anti e1b ap5
    MRE-II RNA interacts in vitro with ORF57 and other cellular proteins. ( A ) Mapping of an MRE binding site for ORF57. Shown on the top is a PAN MRE sequence lacking the MRE-I region in Fig. 4 A, with the 9-nt core of MRE-II loop sequence bolded and underlined and the nt positions of biotinylated RNA oligomers employed in RNA pull-down assays. A cell lysate prepared from TREx BCBL-1-Rta cells 39 induced with Dox for 24 h was used for the RNA pulldown assays with an indicated RNA oligomer. The RNA oligo oNP42 derived from vIL-6 RNA 27 which binds ORF57 was used as a positive control. ORF57 associated with RNA oligos in the pulldowns was immunoblotted using an anti-ORF57 antibody. The cell lysate (10%) before the pulldown was loaded as a Western blot control. ( B ) Biotinylated RNA affinity pulldown analysis by Western blot using anti-ORF57, PABPC1 and <t>E1B-AP5</t> antibodies. Total cell extract from TREx BCBL-1-Rta (R) or -vector (V) cells induced by Dox for 24 h was used for the RNA pulldown assays with each biotinylated RNA oligomer. RNA oligos oNP41 and oNP42 derived from vIL-6 RNA 27 were used, respectively, as a negative and positive oligomer control for ORF57 interaction and 10% of the cell lysate from each cell line before the pulldown was loaded for Western blotting control. RNA oligo oJM68 is a copy of oJM35 with the point mutations in the MRE-II loop described in Fig. 4 B. Control beads indicate no RNA oligomer on the beads.
    Rabbit Polyclonal Anti E1b Ap5, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti e1b ap5/product/Proteintech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti e1b ap5 - by Bioz Stars, 2020-09
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    Proteintech e1b ap5 polyclonal antibody
    MRE-II RNA interacts in vitro with ORF57 and other cellular proteins. ( A ) Mapping of an MRE binding site for ORF57. Shown on the top is a PAN MRE sequence lacking the MRE-I region in Fig. 4 A, with the 9-nt core of MRE-II loop sequence bolded and underlined and the nt positions of biotinylated RNA oligomers employed in RNA pull-down assays. A cell lysate prepared from TREx BCBL-1-Rta cells 39 induced with Dox for 24 h was used for the RNA pulldown assays with an indicated RNA oligomer. The RNA oligo oNP42 derived from vIL-6 RNA 27 which binds ORF57 was used as a positive control. ORF57 associated with RNA oligos in the pulldowns was immunoblotted using an anti-ORF57 antibody. The cell lysate (10%) before the pulldown was loaded as a Western blot control. ( B ) Biotinylated RNA affinity pulldown analysis by Western blot using anti-ORF57, PABPC1 and <t>E1B-AP5</t> antibodies. Total cell extract from TREx BCBL-1-Rta (R) or -vector (V) cells induced by Dox for 24 h was used for the RNA pulldown assays with each biotinylated RNA oligomer. RNA oligos oNP41 and oNP42 derived from vIL-6 RNA 27 were used, respectively, as a negative and positive oligomer control for ORF57 interaction and 10% of the cell lysate from each cell line before the pulldown was loaded for Western blotting control. RNA oligo oJM68 is a copy of oJM35 with the point mutations in the MRE-II loop described in Fig. 4 B. Control beads indicate no RNA oligomer on the beads.
    E1b Ap5 Polyclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e1b ap5 polyclonal antibody/product/Proteintech
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    e1b ap5 polyclonal antibody - by Bioz Stars, 2020-09
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    VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, EIB-AP5 or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using E1B-AP5 antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.

    Journal: EMBO Reports

    Article Title: Vesicular stomatitis virus inhibits mitotic progression and triggers cell death

    doi: 10.1038/embor.2009.179

    Figure Lengend Snippet: VSV M protein interacts with the Rae1–Nup98 complex during both interphase and mitosis. ( A , B ) HeLa cell lysates synchronized at the G1/S boundary and at mitosis were incubated with immobilized recombinant GST–M or GST–M(D) proteins. Bound fractions were analysed by SDS–PAGE, and immunoblot (IB) analysis was carried out with Rae1, Nup98, EIB-AP5 or hnRNP U antibodies. Total lysates were subjected to IB analysis with phospho-histone H3 (Ser 28) antibody. ( C , D ) Mitotic and G1/S lysates were subjected to immunoprecipitation (IP) with Rae1 or Nup98 antibodies in the presence of RNasin or RNase A. Samples were subjected to SDS–PAGE and immunoblot analysis was carried out by using E1B-AP5 antibodies. ( E ) Cells in mitosis were subjected to immunofluorescence with E1B-AP5 and α-tubulin antibodies followed by Apotome microscopy. GST, glutathione- S -transferase; hnRNP, heterogeneous nuclear ribonucleoprotein; M, matrix; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VSV, vesicular stomatitis virus.

    Article Snippet: Blower; ); Rae1 and Nup98 for immunoblot ( ); phospho-histone H3 and Ser 10 (Upstate, USA); β-tubulin–fluorescein isothiocyanate (Sigma, USA); hnRNPA1 (M. Matunis); E1B-AP5 (Proteintech Group, USA); pericentrin (Abcam, USA); monoclonal GST (Santa Cruz Biotechnology, USA); polyclonal γ-tubulin (Sigma); and polyclonal VSV ( ).

    Techniques: Incubation, Recombinant, SDS Page, Immunoprecipitation, Immunofluorescence, Microscopy, Polyacrylamide Gel Electrophoresis

    The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Influenza virus targets the mRNA export machinery and the nuclear pore complex

    doi: 10.1073/pnas.0610977104

    Figure Lengend Snippet: The NS1 protein of influenza virus interacts with key constituents of the mRNA export pathway. ( a ) Cell lysates from 293T cells were incubated with immobilized recombinant GST or GST-NS1. Bound and unbound (UB) fractions were analyzed by 4–20% SDS/PAGE followed by immunoblot analysis with antibodies to NXF1, p15, Rae1, E1B-AP5, and Nup98. ( b and c ) Experiments were performed as in a except that antibodies against Nup96 and against Nup62 and Nup153 (mAb414) were used for immunoblot analysis. ( d ) GST-NS1 or the amino-terminal or carboxyl-terminal domains of NS1 fused with GST were incubated with cell lysates and processed as in a . ( e ) GST-NS1 was incubated with cell lysates untreated or treated with RNase A and processed as in a . ( f and g ) Expression levels of Nups and mRNA export factors in 293T ( f ) and MDCK ( g ) cells infected with influenza virus. Cell extracts were subjected to immunoblot analysis with antibodies against Nup98, β-actin, Rae1, NXF1, and E1B-AP5, and with mAb414 antibodies. ( h ) Half-life measurements of Nup98. MDCK cells were pulse-labeled for 2 h and chased for the depicted time points. Immunoprecipitations were performed with anti-Nup98 antibodies or preimmune serum (PI). Nup98 bands were analyzed by densitometry as described in Methods .

    Article Snippet: Immunoblot analysis was performed with anti-Nup98 , anti-E1B-AP5 (Proteintech Group, Chicago, IL), anti-Rae1 , and anti-NXF1/TAP antibodies (BD Transduction Laboratories, San Jose, CA), anti-p15/NXT antibodies (Abnova, Taipei City, Taiwan), mAb414 , and anti-Influenza A antibodies (virions; Biodesign International, Saco, ME).

    Techniques: Incubation, Recombinant, SDS Page, Expressing, Infection, Labeling

    MRE-II RNA interacts in vitro with ORF57 and other cellular proteins. ( A ) Mapping of an MRE binding site for ORF57. Shown on the top is a PAN MRE sequence lacking the MRE-I region in Fig. 4 A, with the 9-nt core of MRE-II loop sequence bolded and underlined and the nt positions of biotinylated RNA oligomers employed in RNA pull-down assays. A cell lysate prepared from TREx BCBL-1-Rta cells 39 induced with Dox for 24 h was used for the RNA pulldown assays with an indicated RNA oligomer. The RNA oligo oNP42 derived from vIL-6 RNA 27 which binds ORF57 was used as a positive control. ORF57 associated with RNA oligos in the pulldowns was immunoblotted using an anti-ORF57 antibody. The cell lysate (10%) before the pulldown was loaded as a Western blot control. ( B ) Biotinylated RNA affinity pulldown analysis by Western blot using anti-ORF57, PABPC1 and E1B-AP5 antibodies. Total cell extract from TREx BCBL-1-Rta (R) or -vector (V) cells induced by Dox for 24 h was used for the RNA pulldown assays with each biotinylated RNA oligomer. RNA oligos oNP41 and oNP42 derived from vIL-6 RNA 27 were used, respectively, as a negative and positive oligomer control for ORF57 interaction and 10% of the cell lysate from each cell line before the pulldown was loaded for Western blotting control. RNA oligo oJM68 is a copy of oJM35 with the point mutations in the MRE-II loop described in Fig. 4 B. Control beads indicate no RNA oligomer on the beads.

    Journal: International Journal of Biological Sciences

    Article Title: Stability of a Long Noncoding Viral RNA Depends on a 9-nt Core Element at the RNA 5' End to Interact with Viral ORF57 and Cellular PABPC1

    doi:

    Figure Lengend Snippet: MRE-II RNA interacts in vitro with ORF57 and other cellular proteins. ( A ) Mapping of an MRE binding site for ORF57. Shown on the top is a PAN MRE sequence lacking the MRE-I region in Fig. 4 A, with the 9-nt core of MRE-II loop sequence bolded and underlined and the nt positions of biotinylated RNA oligomers employed in RNA pull-down assays. A cell lysate prepared from TREx BCBL-1-Rta cells 39 induced with Dox for 24 h was used for the RNA pulldown assays with an indicated RNA oligomer. The RNA oligo oNP42 derived from vIL-6 RNA 27 which binds ORF57 was used as a positive control. ORF57 associated with RNA oligos in the pulldowns was immunoblotted using an anti-ORF57 antibody. The cell lysate (10%) before the pulldown was loaded as a Western blot control. ( B ) Biotinylated RNA affinity pulldown analysis by Western blot using anti-ORF57, PABPC1 and E1B-AP5 antibodies. Total cell extract from TREx BCBL-1-Rta (R) or -vector (V) cells induced by Dox for 24 h was used for the RNA pulldown assays with each biotinylated RNA oligomer. RNA oligos oNP41 and oNP42 derived from vIL-6 RNA 27 were used, respectively, as a negative and positive oligomer control for ORF57 interaction and 10% of the cell lysate from each cell line before the pulldown was loaded for Western blotting control. RNA oligo oJM68 is a copy of oJM35 with the point mutations in the MRE-II loop described in Fig. 4 B. Control beads indicate no RNA oligomer on the beads.

    Article Snippet: The following antibodies were used in Western blot analyses: a rabbit polyclonal anti-ORF57 antibody (1:3,000 dilution), mouse monoclonal anti-ORF57 antibody (unpublished data, used at a dilution of 1:1,000), rabbit polyclonal anti-PABPC1 (1:700, Abcam ab21060), and rabbit polyclonal anti-E1B-AP5 (1:1,000, ProteinTech Group 0578-1-AP, Chicago, IL), together with corresponding peroxidase-conjugated secondary antibodies (1:10,000, Sigma).

    Techniques: In Vitro, Binding Assay, Sequencing, Derivative Assay, Positive Control, Western Blot, Plasmid Preparation