oligo resuspension buffer Search Results


oligo resuspension buffer  (Thermo Fisher)
99
Thermo Fisher oligo resuspension buffer
Oligo Resuspension Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pronto buffer  (Corning Life Sciences)
90
Corning Life Sciences pronto buffer
Pronto Buffer, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
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rna ligation mix  (New England Biolabs)
86
New England Biolabs rna ligation mix
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Rna Ligation Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rna ligation mix/product/New England Biolabs
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100-μl nucleofector solution v  (Amaxa)
90
Amaxa 100-μl nucleofector solution v
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
100 μl Nucleofector Solution V, supplied by Amaxa, 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/100-μl nucleofector solution v/product/Amaxa
Average 90 stars, based on 1 article reviews
100-μl nucleofector solution v - by Bioz Stars, 2026-03
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oligo dt wash buffer  (Bio-Rad)
98
Bio-Rad oligo dt wash buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Oligo Dt Wash Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/oligo dt wash buffer/product/Bio-Rad
Average 98 stars, based on 1 article reviews
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oligo hyb buffer  (Full Moon BioSystems)
90
Full Moon BioSystems oligo hyb buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Oligo Hyb Buffer, supplied by Full Moon BioSystems, 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/oligo hyb buffer/product/Full Moon BioSystems
Average 90 stars, based on 1 article reviews
oligo hyb buffer - by Bioz Stars, 2026-03
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ultrahyb oligo hybridization buffer  (Thermo Fisher)
90
Thermo Fisher ultrahyb oligo hybridization buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Ultrahyb Oligo Hybridization Buffer, supplied by Thermo Fisher, 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/ultrahyb oligo hybridization buffer/product/Thermo Fisher
Average 90 stars, based on 1 article reviews
ultrahyb oligo hybridization buffer - by Bioz Stars, 2026-03
90/100 stars
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duplex buffer  (Danaher Inc)
86
Danaher Inc duplex buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Duplex Buffer, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/duplex buffer/product/Danaher Inc
Average 86 stars, based on 1 article reviews
duplex buffer - by Bioz Stars, 2026-03
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zymolase buffer  (Bio-Rad)
96
Bio-Rad zymolase buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Zymolase Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/zymolase buffer/product/Bio-Rad
Average 96 stars, based on 1 article reviews
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1x annealing buffer  (Thermo Fisher)
90
Thermo Fisher 1x annealing buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
1x Annealing Buffer, supplied by Thermo Fisher, 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/1x annealing buffer/product/Thermo Fisher
Average 90 stars, based on 1 article reviews
1x annealing buffer - by Bioz Stars, 2026-03
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long oligo hybridization solution  (Corning Life Sciences)
90
Corning Life Sciences long oligo hybridization solution
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Long Oligo Hybridization Solution, supplied by Corning Life Sciences, 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/long oligo hybridization solution/product/Corning Life Sciences
Average 90 stars, based on 1 article reviews
long oligo hybridization solution - by Bioz Stars, 2026-03
90/100 stars
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nexterion oligo hyb buffer  (PEQLAB)
90
PEQLAB nexterion oligo hyb buffer
(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize <t>RNA-protein</t> complexes. The protein of interest is stringently immunoprecipitated <t>and</t> <t>UV-crosslinked</t> RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.
Nexterion Oligo Hyb Buffer, supplied by PEQLAB, 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/nexterion oligo hyb buffer/product/PEQLAB
Average 90 stars, based on 1 article reviews
nexterion oligo hyb buffer - by Bioz Stars, 2026-03
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Image Search Results


(A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize RNA-protein complexes. The protein of interest is stringently immunoprecipitated and UV-crosslinked RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.

Journal: bioRxiv

Article Title: Cellular RNA Interacts with MAVS to Promote Antiviral Signaling

doi: 10.1101/2023.09.25.559083

Figure Lengend Snippet: (A) Infrared-dye crosslinking and immunoprecipitation (irCLIP) strategy to visualize RNA-protein complexes. The protein of interest is stringently immunoprecipitated and UV-crosslinked RNA is digested to fragments with RNase A. Following ligation of an IRDye-800 conjugated oligonucleotide, complexes are resolved by SDS-PAGE. RNA-protein complexes are detected by IRDye-800 fluorescence and immunoprecipitation is validated by immunoblot analysis. (B) irCLIP −/+ crosslinking of FLAG-tagged MAVS expressed in 293T MAVS KO cells (24 hpt). EV = empty vector. (C) Quantification of IRDye signal in experiments in (B). (D) irCLIP of endogenous MAVS from mock- and SenV-infected (100 HAU/mL, 16 hpi) 293T cells. (E) Quantification of IRDye signal in experiments in (D). (F) irCLIP of FLAG-tagged murine MAVS expressed in murine NIH3T3 cells (24 hpt). (G) Quantification of IRDye signal in experiments in (F). (H) Schematic of FLAG-tagged full length MAVS and MAVSΔ103-467 used in (I). (I) irCLIP of the indicated FLAG-tagged MAVS constructs expressed in 293T MAVS KO cells (24 hpt). (J) Quantification of IRDye signal in experiments in (I). (K) Prediction of disorder in human MAVS (red) and in 325 mammalian species (blue) using IUPred3. Data in (B), Data in (D), (F), and (I) are representative of 3 biological replicates. Values in (C), (E), and (G), and (J) are mean ± SEM of 3 biological replicates. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 by one-way ANOVA with Tukey’s multiple comparison test. n.s. = not significant.

Article Snippet: For ligation of IRDye-800 conjugated oligo to RNA crosslinked to protein, beads were resuspended in 30 µL RNA ligation mix (1X RNA ligase I buffer (NEB), 1 µL RNA ligase I (NEB), 0.5 µL IRDye800-labeled oligonucleotide , 5 µL PEG400, and 0.5 µL RNaseIN) and incubated for 16 hrs at 16°C with shaking in Thermomixer (1200 rpm).

Techniques: Immunoprecipitation, Ligation, SDS Page, Fluorescence, Western Blot, Plasmid Preparation, Infection, Construct, Comparison