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InvivoGen superscript iii reverse transcriptase
Activation deficiency of chicken and porcine TLR5 can be partially rescued with respective TLR5 chimeras. HeLa ( A ) or HEK293-T cells ( B ) were transiently transfected with adjusted amounts of plasmid <t>DNA</t> encoding for TLR5 from different species or chimeric receptors (transfected plasmid amounts see Fig. 1 ). 48 h after transfection, cells were coincubated with purified recombinant Salmonella FliC (50 ng/well) or mock-treated for four hours. IL-8 secretion in the cell supernatants was determined by ELISA. IL-8 secretion of hTLR5-transfected, FliC-stimulated cells was set to 100% (reference); relative IL-8 secretion of all constructs with regard to the reference is depicted. Each condition was tested in two independent biological replicates (each in technical triplicates), the results of which are summarized here as mean and standard error. One representative experiment out of <t>three</t> is shown. For all constructs in A and B, except for pEF6-empty, the differences between mock-coincubated and FliC-coincubated condition were highly significant (Student’s t -test, unpaired, two-tailed; p
Superscript Iii Reverse Transcriptase, supplied by InvivoGen, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition

Journal: Scientific Reports

doi: 10.1038/s41598-018-29371-0

Activation deficiency of chicken and porcine TLR5 can be partially rescued with respective TLR5 chimeras. HeLa ( A ) or HEK293-T cells ( B ) were transiently transfected with adjusted amounts of plasmid DNA encoding for TLR5 from different species or chimeric receptors (transfected plasmid amounts see Fig. 1 ). 48 h after transfection, cells were coincubated with purified recombinant Salmonella FliC (50 ng/well) or mock-treated for four hours. IL-8 secretion in the cell supernatants was determined by ELISA. IL-8 secretion of hTLR5-transfected, FliC-stimulated cells was set to 100% (reference); relative IL-8 secretion of all constructs with regard to the reference is depicted. Each condition was tested in two independent biological replicates (each in technical triplicates), the results of which are summarized here as mean and standard error. One representative experiment out of three is shown. For all constructs in A and B, except for pEF6-empty, the differences between mock-coincubated and FliC-coincubated condition were highly significant (Student’s t -test, unpaired, two-tailed; p
Figure Legend Snippet: Activation deficiency of chicken and porcine TLR5 can be partially rescued with respective TLR5 chimeras. HeLa ( A ) or HEK293-T cells ( B ) were transiently transfected with adjusted amounts of plasmid DNA encoding for TLR5 from different species or chimeric receptors (transfected plasmid amounts see Fig. 1 ). 48 h after transfection, cells were coincubated with purified recombinant Salmonella FliC (50 ng/well) or mock-treated for four hours. IL-8 secretion in the cell supernatants was determined by ELISA. IL-8 secretion of hTLR5-transfected, FliC-stimulated cells was set to 100% (reference); relative IL-8 secretion of all constructs with regard to the reference is depicted. Each condition was tested in two independent biological replicates (each in technical triplicates), the results of which are summarized here as mean and standard error. One representative experiment out of three is shown. For all constructs in A and B, except for pEF6-empty, the differences between mock-coincubated and FliC-coincubated condition were highly significant (Student’s t -test, unpaired, two-tailed; p

Techniques Used: Activation Assay, Transfection, Plasmid Preparation, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Construct, Two Tailed Test

Related Articles

SYBR Green Assay:

Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition
Article Snippet: 1 µg of total DNA-free RNA was used for cDNA synthesis with Superscript III reverse transcriptase (Invivogen) and oligo(dT) primers (Invivogen). .. 1 µl of synthesized cDNA was used for quantitative RT-PCR based on SYBR green (Qiagen) chemistry.

Isolation:

Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition
Article Snippet: DNAse I treatment of the isolated RNA was performed with Turbo DNA-free Kit (Ambion) according to manufacturer’s instructions. .. 1 µg of total DNA-free RNA was used for cDNA synthesis with Superscript III reverse transcriptase (Invivogen) and oligo(dT) primers (Invivogen).

Quantitative RT-PCR:

Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition
Article Snippet: Paragraph title: RNA preparation and quantitative real-time (RT) PCR ... 1 µg of total DNA-free RNA was used for cDNA synthesis with Superscript III reverse transcriptase (Invivogen) and oligo(dT) primers (Invivogen).

Expressing:

Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition
Article Snippet: 1 µg of total DNA-free RNA was used for cDNA synthesis with Superscript III reverse transcriptase (Invivogen) and oligo(dT) primers (Invivogen). .. Normalisation of the results was carried out according to transcriptional expression of chicken glyceraldehyde-3-phosphate dehydrogenase (chGAPDH).

Synthesized:

Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition
Article Snippet: 1 µg of total DNA-free RNA was used for cDNA synthesis with Superscript III reverse transcriptase (Invivogen) and oligo(dT) primers (Invivogen). .. 1 µl of synthesized cDNA was used for quantitative RT-PCR based on SYBR green (Qiagen) chemistry.

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    InvivoGen superscript iii reverse transcriptase
    Activation deficiency of chicken and porcine TLR5 can be partially rescued with respective TLR5 chimeras. HeLa ( A ) or HEK293-T cells ( B ) were transiently transfected with adjusted amounts of plasmid <t>DNA</t> encoding for TLR5 from different species or chimeric receptors (transfected plasmid amounts see Fig. 1 ). 48 h after transfection, cells were coincubated with purified recombinant Salmonella FliC (50 ng/well) or mock-treated for four hours. IL-8 secretion in the cell supernatants was determined by ELISA. IL-8 secretion of hTLR5-transfected, FliC-stimulated cells was set to 100% (reference); relative IL-8 secretion of all constructs with regard to the reference is depicted. Each condition was tested in two independent biological replicates (each in technical triplicates), the results of which are summarized here as mean and standard error. One representative experiment out of <t>three</t> is shown. For all constructs in A and B, except for pEF6-empty, the differences between mock-coincubated and FliC-coincubated condition were highly significant (Student’s t -test, unpaired, two-tailed; p
    Superscript Iii Reverse Transcriptase, supplied by InvivoGen, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii reverse transcriptase/product/InvivoGen
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript iii reverse transcriptase - by Bioz Stars, 2020-04
    93/100 stars
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    Activation deficiency of chicken and porcine TLR5 can be partially rescued with respective TLR5 chimeras. HeLa ( A ) or HEK293-T cells ( B ) were transiently transfected with adjusted amounts of plasmid DNA encoding for TLR5 from different species or chimeric receptors (transfected plasmid amounts see Fig. 1 ). 48 h after transfection, cells were coincubated with purified recombinant Salmonella FliC (50 ng/well) or mock-treated for four hours. IL-8 secretion in the cell supernatants was determined by ELISA. IL-8 secretion of hTLR5-transfected, FliC-stimulated cells was set to 100% (reference); relative IL-8 secretion of all constructs with regard to the reference is depicted. Each condition was tested in two independent biological replicates (each in technical triplicates), the results of which are summarized here as mean and standard error. One representative experiment out of three is shown. For all constructs in A and B, except for pEF6-empty, the differences between mock-coincubated and FliC-coincubated condition were highly significant (Student’s t -test, unpaired, two-tailed; p

    Journal: Scientific Reports

    Article Title: Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition

    doi: 10.1038/s41598-018-29371-0

    Figure Lengend Snippet: Activation deficiency of chicken and porcine TLR5 can be partially rescued with respective TLR5 chimeras. HeLa ( A ) or HEK293-T cells ( B ) were transiently transfected with adjusted amounts of plasmid DNA encoding for TLR5 from different species or chimeric receptors (transfected plasmid amounts see Fig. 1 ). 48 h after transfection, cells were coincubated with purified recombinant Salmonella FliC (50 ng/well) or mock-treated for four hours. IL-8 secretion in the cell supernatants was determined by ELISA. IL-8 secretion of hTLR5-transfected, FliC-stimulated cells was set to 100% (reference); relative IL-8 secretion of all constructs with regard to the reference is depicted. Each condition was tested in two independent biological replicates (each in technical triplicates), the results of which are summarized here as mean and standard error. One representative experiment out of three is shown. For all constructs in A and B, except for pEF6-empty, the differences between mock-coincubated and FliC-coincubated condition were highly significant (Student’s t -test, unpaired, two-tailed; p

    Article Snippet: 1 µg of total DNA-free RNA was used for cDNA synthesis with Superscript III reverse transcriptase (Invivogen) and oligo(dT) primers (Invivogen).

    Techniques: Activation Assay, Transfection, Plasmid Preparation, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Construct, Two Tailed Test