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  • 99
    Thermo Fisher antibody mixture anti mouse poly hpo
    Antibody Mixture Anti Mouse Poly Hpo, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody mixture anti mouse poly hpo/product/Thermo Fisher
    Average 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    antibody mixture anti mouse poly hpo - by Bioz Stars, 2020-11
    99/100 stars
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    99
    Millipore sodium phosphate dibasic
    Sodium Phosphate Dibasic, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 824 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sodium phosphate dibasic/product/Millipore
    Average 99 stars, based on 824 article reviews
    Price from $9.99 to $1999.99
    sodium phosphate dibasic - by Bioz Stars, 2020-11
    99/100 stars
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    91
    Carl Zeiss zeiss hpo
    Zeiss Hpo, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 91/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/zeiss hpo/product/Carl Zeiss
    Average 91 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    zeiss hpo - by Bioz Stars, 2020-11
    91/100 stars
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    90
    The Jackson Laboratory hpo gene annotations
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Hpo Gene Annotations, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hpo gene annotations/product/The Jackson Laboratory
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hpo gene annotations - by Bioz Stars, 2020-11
    90/100 stars
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    hpo  (MedGen)
    92
    MedGen hpo
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Hpo, supplied by MedGen, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hpo/product/MedGen
    Average 92 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    hpo - by Bioz Stars, 2020-11
    92/100 stars
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    85
    World Precision Instruments iso hpo 2 electrode
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Iso Hpo 2 Electrode, supplied by World Precision Instruments, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iso hpo 2 electrode/product/World Precision Instruments
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    iso hpo 2 electrode - by Bioz Stars, 2020-11
    85/100 stars
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    92
    Cell Signaling Technology Inc anti phospho hpo
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Anti Phospho Hpo, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho hpo/product/Cell Signaling Technology Inc
    Average 92 stars, based on 18 article reviews
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    anti phospho hpo - by Bioz Stars, 2020-11
    92/100 stars
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    86
    Cell Signaling Technology Inc goat anti rabbit hpo
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Goat Anti Rabbit Hpo, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit hpo/product/Cell Signaling Technology Inc
    Average 86 stars, based on 16 article reviews
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    goat anti rabbit hpo - by Bioz Stars, 2020-11
    86/100 stars
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    85
    Ashcroft Inc heise model hpo pressure transducer
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Heise Model Hpo Pressure Transducer, supplied by Ashcroft Inc, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/heise model hpo pressure transducer/product/Ashcroft Inc
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    heise model hpo pressure transducer - by Bioz Stars, 2020-11
    85/100 stars
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    86
    Bio-Rad hpo color development kit
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Hpo Color Development Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hpo color development kit/product/Bio-Rad
    Average 86 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    hpo color development kit - by Bioz Stars, 2020-11
    86/100 stars
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    91
    The Jackson Laboratory hpo terms
    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of <t>HPO</t> terms, which act as input for Phen2Gene or <t>Phenolyzer.</t> These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.
    Hpo Terms, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hpo terms/product/The Jackson Laboratory
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hpo terms - by Bioz Stars, 2020-11
    91/100 stars
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    85
    Santa Cruz Biotechnology anti rabbit igg hpo
    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit <t>IgG/HPO</t> (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
    Anti Rabbit Igg Hpo, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit igg hpo/product/Santa Cruz Biotechnology
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    anti rabbit igg hpo - by Bioz Stars, 2020-11
    85/100 stars
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    90
    SeraCare anti swine igg conjugated to hpo
    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit <t>IgG/HPO</t> (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
    Anti Swine Igg Conjugated To Hpo, supplied by SeraCare, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti swine igg conjugated to hpo/product/SeraCare
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti swine igg conjugated to hpo - by Bioz Stars, 2020-11
    90/100 stars
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    86
    Thermo Fisher goat anti rabbit poly hpo
    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit <t>IgG/HPO</t> (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
    Goat Anti Rabbit Poly Hpo, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit poly hpo/product/Thermo Fisher
    Average 86 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit poly hpo - by Bioz Stars, 2020-11
    86/100 stars
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    99
    Millipore disodium hydrogen phosphate na 2 hpo 4
    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit <t>IgG/HPO</t> (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
    Disodium Hydrogen Phosphate Na 2 Hpo 4, supplied by Millipore, 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/disodium hydrogen phosphate na 2 hpo 4/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    disodium hydrogen phosphate na 2 hpo 4 - by Bioz Stars, 2020-11
    99/100 stars
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    90
    Clinithink clix focus quality controlclix focus extracts hpo terms
    RC variant annotation workflow using Emedgene. Flowchart workflow for evaluating variants prioritized by Emedgene using manually curated <t>HPO</t> terms, as well as <t>CLiX</t> Focus-derived HPO terms.
    Clix Focus Quality Controlclix Focus Extracts Hpo Terms, supplied by Clinithink, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/clix focus quality controlclix focus extracts hpo terms/product/Clinithink
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    clix focus quality controlclix focus extracts hpo terms - by Bioz Stars, 2020-11
    90/100 stars
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    99
    Millipore peanut oil hpo
    RC variant annotation workflow using Emedgene. Flowchart workflow for evaluating variants prioritized by Emedgene using manually curated <t>HPO</t> terms, as well as <t>CLiX</t> Focus-derived HPO terms.
    Peanut Oil Hpo, supplied by Millipore, 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/peanut oil hpo/product/Millipore
    Average 99 stars, based on 1 article reviews
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    peanut oil hpo - by Bioz Stars, 2020-11
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    85
    Agilent technologies horseradish peroxidase hpo conjugated anti ratigg
    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated <t>anti-RatIgG</t> (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
    Horseradish Peroxidase Hpo Conjugated Anti Ratigg, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/horseradish peroxidase hpo conjugated anti ratigg/product/Agilent technologies
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    horseradish peroxidase hpo conjugated anti ratigg - by Bioz Stars, 2020-11
    85/100 stars
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    85
    Medicago hydroperoxide hpo lyase
    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated <t>anti-RatIgG</t> (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
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    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase <t>(HPO)</t> conjugated <t>anti-RatIgG</t> (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.
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    Par-1 disrupts the association of the <t>Hpo-Sav</t> complex in a kinase-dependent manner. (A) Par-1 could destabilize Hpo-induced accumulation of Sav in a kinase-dependent manner. S2 cells were transfected with the indicated constructs followed by a Western blot analysis. CFP served as a loading control. (B) Par-1 inhibits the phosphorylation of Sav induced by Hpo. The mobility shift assay was employed. The loading volume was adjusted according to the total Sav protein level. (C) Par-1 disrupts the interaction of the Hpo-Sav complex. S2 cells were transfected with the indicated constructs followed by <t>immunoprecipitation</t> to test whether the interaction between Sav and endogenous Hpo was affected by Par-1. Note that less Sav interacted with Hpo in the presence of Par-1. (D–D″) Par-1 RNAi is incapable of inhibiting sav mutant-induced adult eye overgrowth. The clones were generated using the MARCM system. The genotypes are as following: eyflp, ubi-Gal4, UAS-GFP; FRT82B/FRT82BGal80 (D), e yflp, ubi-Gal4, UAS-GFP; FRT82B Sav SH13 / FRT82B Gal80 (D′), and eyflp, ubi-Gal4, UAS-GFP; Par-1-RNAi; FRT82B Sav SH13 / FRT82B Gal80 (D″). (E) The proposed mechanism of Par-1 regulation of the Hpo pathway (see text for further detail).
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    Hppy and Msn promote phosphorylation of Wts but not <t>Hpo.</t> ( a , b ) Western blot analysis of cell lysates derived from S2 cells expressing the indicated constructs. Hppy, Msn, as well as Hpo stimulated phosphorylation of Wts at T1077, which was recognized by a phospho-specific antibody p-Wts ( a ). By contrast, neither Hppy nor Msn stimulated phosphorylation of Hpo at T195 recognized by a phospho-specific antibody <t>p-Hpo/MST1/2</t> ( b ). ( c ) Hppy formed a complex with Wts in S2 cells as revealed by a co-immunoprecipitation assay.
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    Hppy and Msn promote phosphorylation of Wts but not <t>Hpo.</t> ( a , b ) Western blot analysis of cell lysates derived from S2 cells expressing the indicated constructs. Hppy, Msn, as well as Hpo stimulated phosphorylation of Wts at T1077, which was recognized by a phospho-specific antibody p-Wts ( a ). By contrast, neither Hppy nor Msn stimulated phosphorylation of Hpo at T195 recognized by a phospho-specific antibody <t>p-Hpo/MST1/2</t> ( b ). ( c ) Hppy formed a complex with Wts in S2 cells as revealed by a co-immunoprecipitation assay.
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    Hppy and Msn promote phosphorylation of Wts but not <t>Hpo.</t> ( a , b ) Western blot analysis of cell lysates derived from S2 cells expressing the indicated constructs. Hppy, Msn, as well as Hpo stimulated phosphorylation of Wts at T1077, which was recognized by a phospho-specific antibody p-Wts ( a ). By contrast, neither Hppy nor Msn stimulated phosphorylation of Hpo at T195 recognized by a phospho-specific antibody <t>p-Hpo/MST1/2</t> ( b ). ( c ) Hppy formed a complex with Wts in S2 cells as revealed by a co-immunoprecipitation assay.
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    Hppy and Msn promote phosphorylation of Wts but not <t>Hpo.</t> ( a , b ) Western blot analysis of cell lysates derived from S2 cells expressing the indicated constructs. Hppy, Msn, as well as Hpo stimulated phosphorylation of Wts at T1077, which was recognized by a phospho-specific antibody p-Wts ( a ). By contrast, neither Hppy nor Msn stimulated phosphorylation of Hpo at T195 recognized by a phospho-specific antibody <t>p-Hpo/MST1/2</t> ( b ). ( c ) Hppy formed a complex with Wts in S2 cells as revealed by a co-immunoprecipitation assay.
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    Image Search Results


    General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of HPO terms, which act as input for Phen2Gene or Phenolyzer. These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.

    Journal: bioRxiv

    Article Title: Phen2Gene: Rapid Phenotype-Driven Gene Prioritization for Rare Diseases

    doi: 10.1101/870527

    Figure Lengend Snippet: General use case. Proband has a condition with unknown genetic cause but several candidate variants annotated and filtered using ANNOVAR 104 . Clinical notes on the proband’s condition are used by Doc2HPO to generate a list of HPO terms, which act as input for Phen2Gene or Phenolyzer. These tools rank several thousand genes and by intersecting them with the candidate list of genes overlapping the variants, we obtain a list of likely candidate genes for KGB syndrome, which is known to be caused by variants in ANKRD11, shown here.

    Article Snippet: We updated the databases inside Phenolyzer, incorporated new HPO-gene annotations from the Jackson Laboratory database, fixed some bugs, and released it as Enhanced Phenolyzer (ver.

    Techniques:

    The construction of the HPO2Gene Knowledgebase. HPO terms are extracted one by one from the HPO database and passed into an enhanced version of Phenolyzer (dubbed Enhanced Phenolyzer) to create a database of ranked gene lists for all HPO terms. (a) The workflow of Enhanced Phenolyzer. (b) Construction of the HPO2Gene Knowledgebase.

    Journal: bioRxiv

    Article Title: Phen2Gene: Rapid Phenotype-Driven Gene Prioritization for Rare Diseases

    doi: 10.1101/870527

    Figure Lengend Snippet: The construction of the HPO2Gene Knowledgebase. HPO terms are extracted one by one from the HPO database and passed into an enhanced version of Phenolyzer (dubbed Enhanced Phenolyzer) to create a database of ranked gene lists for all HPO terms. (a) The workflow of Enhanced Phenolyzer. (b) Construction of the HPO2Gene Knowledgebase.

    Article Snippet: We updated the databases inside Phenolyzer, incorporated new HPO-gene annotations from the Jackson Laboratory database, fixed some bugs, and released it as Enhanced Phenolyzer (ver.

    Techniques:

    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase (HPO) conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.

    Journal: Virology Journal

    Article Title: The effects of N-terminal insertion into VSV-G of an scFv peptide

    doi: 10.1186/1743-422X-3-69

    Figure Lengend Snippet: scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase (HPO) conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.

    Article Snippet: VSV-G and scFv/VSV-G carry an HA tag, and were detected by a rat anti-HA antibody (Sigma), followed by a horseradish peroxidase (HPO) conjugated anti-RatIgG (Dako). p24Gag, detected by SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) and an anti-rabbit IgG/HPO (Santa Cruz), was used as an internal reference to normalise the virion proteins.

    Techniques: Expressing, Derivative Assay, Plasmid Preparation, Polymerase Chain Reaction, Clone Assay, Sequencing, Immunodetection, Construct, Generated, Amplification, Modification, Produced, Transfection

    RC variant annotation workflow using Emedgene. Flowchart workflow for evaluating variants prioritized by Emedgene using manually curated HPO terms, as well as CLiX Focus-derived HPO terms.

    Journal: NPJ Genomic Medicine

    Article Title: Children’s rare disease cohorts: an integrative research and clinical genomics initiative

    doi: 10.1038/s41525-020-0137-0

    Figure Lengend Snippet: RC variant annotation workflow using Emedgene. Flowchart workflow for evaluating variants prioritized by Emedgene using manually curated HPO terms, as well as CLiX Focus-derived HPO terms.

    Article Snippet: Clinithink CLiX Focus quality controlCLiX Focus extracts HPO terms on a per note basis and provides a note-frequency ranked list of HPO terms for each patient.

    Techniques: Variant Assay, Derivative Assay

    scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase (HPO) conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.

    Journal: Virology Journal

    Article Title: The effects of N-terminal insertion into VSV-G of an scFv peptide

    doi: 10.1186/1743-422X-3-69

    Figure Lengend Snippet: scFv-VSV-G expression plasmids and incorporation in HIV-1 derived particles. a) Expression plasmids: (I) VSV-G expression plasmid (PM 730). The 1.6 kB HindIII-BamHI VSV-G fragment (serotype Indiana) was transferred from pFB.VSVG (J.M. Heard, Paris, France) into pcDNA3 (InVitrogen) by PCR cloning according to standard procedures. A haemagglutinin (HA) sequence was added at the C terminus of VSV-G for immunodetection. (II and III) scFv/VSV-G expression plasmids. The chimaeric constructs were generated by PCR-based cloning. Mature VSV-G (from amino acid 17) was amplified from PM 730 and introduced into the PM 441 and PM 442 plasmids [23]. These constructs originate from an MLV-derived plasmid (FBMOSALF [31]), modified to contain a scFv (αMHC and αHEL, respectively [27]), upstream of the GP gene. Consequently, the resulting constructs (αMHC/VSV-G and αHEL/VSV-G) express the genes from the MLV LTR, with a MLV leader sequence (L mlv ) and 6 additional amino acids from the virus. (IV and V) Vectors for production of HIV-1 derived viral particles. A HIV-1-based lentiviral vector (V) (CD 416; pHRCMV) [29], into which EGFP gene had been inserted, together with a helper plasmid (CD 417; pCMV△R8.2) [29] expressing Gag, Pol and accessory HIV-1 proteins (IV), were used for production of HIV-1 particles. Expression vectors, physical maps and primer sequences are available upon request. b. VSV-G- and scFv/VSV-G pseudotyped HIV-1 particles, produced in 293T cells. 2 × 10 6 293T cells in 10 cm diameter tissue culture dishes were transiently transfected with 5 μg of an LTR-driven EGFP vector (pHRCMV-EGFP) and 4 μg of a helper plasmid (pCMV△R8.2) [29], using the calcium phosphate co-precipitation procedure [36]. 5 μg PM 730 or 30 μg scFv/VSV-G plasmid (PM 981 or PM 983) were also included. DNA precipitates were removed after 16 hours, and the viral supernatants were collected 24–48 hours later and pelleted by ultracentrifugation (BeckmanCoulter) at 25 kRPM, 4°C for 2 hours and resuspended in 1% of the original volume. Cell lysates and 2 μl concentrated scFv/VSV-G virus or 10 μl of non-concentrated VSV-G virus were separated on a 12 % SDS polyacrylamide gel and transferred onto Protran nitrocellulose membranes (Schleicher and Schuell). HA-tagged VSV-G and scFv/VSV-G were detected using a rat anti-HA antibody (Sigma), followed by horse radish peroxidase (HPO) conjugated anti-RatIgG (Dako). p24Gag was detected using SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) followed by anti-rabbit IgG/HPO (Santa Cruz), and was used as an internal reference to normalise for the virion protein quantities. The membranes were developed with Renaissance chemoluminescence kit (NEN Life Science Products), as recommended by the supplier.

    Article Snippet: VSV-G and scFv/VSV-G carry an HA tag, and were detected by a rat anti-HA antibody (Sigma), followed by a horseradish peroxidase (HPO) conjugated anti-RatIgG (Dako). p24Gag, detected by SF2 rabbit monoclonal antibody (NIH AIDS Research and Reference Reagent Program) and an anti-rabbit IgG/HPO (Santa Cruz), was used as an internal reference to normalise the virion proteins.

    Techniques: Expressing, Derivative Assay, Plasmid Preparation, Polymerase Chain Reaction, Clone Assay, Sequencing, Immunodetection, Construct, Generated, Amplification, Modification, Produced, Transfection

    Par-1 disrupts the association of the Hpo-Sav complex in a kinase-dependent manner. (A) Par-1 could destabilize Hpo-induced accumulation of Sav in a kinase-dependent manner. S2 cells were transfected with the indicated constructs followed by a Western blot analysis. CFP served as a loading control. (B) Par-1 inhibits the phosphorylation of Sav induced by Hpo. The mobility shift assay was employed. The loading volume was adjusted according to the total Sav protein level. (C) Par-1 disrupts the interaction of the Hpo-Sav complex. S2 cells were transfected with the indicated constructs followed by immunoprecipitation to test whether the interaction between Sav and endogenous Hpo was affected by Par-1. Note that less Sav interacted with Hpo in the presence of Par-1. (D–D″) Par-1 RNAi is incapable of inhibiting sav mutant-induced adult eye overgrowth. The clones were generated using the MARCM system. The genotypes are as following: eyflp, ubi-Gal4, UAS-GFP; FRT82B/FRT82BGal80 (D), e yflp, ubi-Gal4, UAS-GFP; FRT82B Sav SH13 / FRT82B Gal80 (D′), and eyflp, ubi-Gal4, UAS-GFP; Par-1-RNAi; FRT82B Sav SH13 / FRT82B Gal80 (D″). (E) The proposed mechanism of Par-1 regulation of the Hpo pathway (see text for further detail).

    Journal: PLoS Biology

    Article Title: Par-1 Regulates Tissue Growth by Influencing Hippo Phosphorylation Status and Hippo-Salvador Association

    doi: 10.1371/journal.pbio.1001620

    Figure Lengend Snippet: Par-1 disrupts the association of the Hpo-Sav complex in a kinase-dependent manner. (A) Par-1 could destabilize Hpo-induced accumulation of Sav in a kinase-dependent manner. S2 cells were transfected with the indicated constructs followed by a Western blot analysis. CFP served as a loading control. (B) Par-1 inhibits the phosphorylation of Sav induced by Hpo. The mobility shift assay was employed. The loading volume was adjusted according to the total Sav protein level. (C) Par-1 disrupts the interaction of the Hpo-Sav complex. S2 cells were transfected with the indicated constructs followed by immunoprecipitation to test whether the interaction between Sav and endogenous Hpo was affected by Par-1. Note that less Sav interacted with Hpo in the presence of Par-1. (D–D″) Par-1 RNAi is incapable of inhibiting sav mutant-induced adult eye overgrowth. The clones were generated using the MARCM system. The genotypes are as following: eyflp, ubi-Gal4, UAS-GFP; FRT82B/FRT82BGal80 (D), e yflp, ubi-Gal4, UAS-GFP; FRT82B Sav SH13 / FRT82B Gal80 (D′), and eyflp, ubi-Gal4, UAS-GFP; Par-1-RNAi; FRT82B Sav SH13 / FRT82B Gal80 (D″). (E) The proposed mechanism of Par-1 regulation of the Hpo pathway (see text for further detail).

    Article Snippet: The following antibodies were used in the immunoprecipitation or Western blot analyses: rabbit anti-Hpo antibody , rabbit anti-Phospho Hpo (Thr195) antibody (Cell Signaling Technology), mouse anti-Flag antibody (Sigma), mouse anti-Myc antibody (Santa Cruz), mouse anti-V5 antibody (Invitrogen), and mouse anti-GFP/CFP antibody (Santa Cruz).

    Techniques: Transfection, Construct, Western Blot, Mobility Shift, Immunoprecipitation, Mutagenesis, Clone Assay, Generated

    Hppy and Msn promote phosphorylation of Wts but not Hpo. ( a , b ) Western blot analysis of cell lysates derived from S2 cells expressing the indicated constructs. Hppy, Msn, as well as Hpo stimulated phosphorylation of Wts at T1077, which was recognized by a phospho-specific antibody p-Wts ( a ). By contrast, neither Hppy nor Msn stimulated phosphorylation of Hpo at T195 recognized by a phospho-specific antibody p-Hpo/MST1/2 ( b ). ( c ) Hppy formed a complex with Wts in S2 cells as revealed by a co-immunoprecipitation assay.

    Journal: Cell Discovery

    Article Title: Overlapping functions of the MAP4K family kinases Hppy and Msn in Hippo signaling

    doi: 10.1038/celldisc.2015.38

    Figure Lengend Snippet: Hppy and Msn promote phosphorylation of Wts but not Hpo. ( a , b ) Western blot analysis of cell lysates derived from S2 cells expressing the indicated constructs. Hppy, Msn, as well as Hpo stimulated phosphorylation of Wts at T1077, which was recognized by a phospho-specific antibody p-Wts ( a ). By contrast, neither Hppy nor Msn stimulated phosphorylation of Hpo at T195 recognized by a phospho-specific antibody p-Hpo/MST1/2 ( b ). ( c ) Hppy formed a complex with Wts in S2 cells as revealed by a co-immunoprecipitation assay.

    Article Snippet: Antibodies used were as follows: mouse anti-Myc (Santa Cruz Biotechnology, Santa Cruz, CA, USA, sc-40); mouse anti-HA (Santa Cruz Biotechnology, sc-7392); mouse anti-Flag (Sigma, M2 F3165); rabbit anti-p-Hpo/Mst1/2 (Cell Signaling Technology, Danvers, MA, USA, #3681).

    Techniques: Western Blot, Derivative Assay, Expressing, Construct, Co-Immunoprecipitation Assay