mouse anti rat igg  (Vector Laboratories)


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    Vector Laboratories mouse anti rat igg
    Mouse Anti Rat Igg, supplied by Vector Laboratories, 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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    mouse anti rat igg  (Vector Laboratories)


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    Vector Laboratories mouse anti rat igg
    Mouse Anti Rat Igg, supplied by Vector Laboratories, 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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    hrp conjugated goat anti mouse secondary antibody  (Vector Laboratories)


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    Vector Laboratories hrp conjugated goat anti mouse secondary antibody
    Hrp Conjugated Goat Anti Mouse Secondary Antibody, supplied by Vector Laboratories, 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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    alexa fluor 568 goat anti rat igg intritrogen  (Vector Laboratories)


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    Vector Laboratories alexa fluor 568 goat anti rat igg intritrogen
    Alexa Fluor 568 Goat Anti Rat Igg Intritrogen, supplied by Vector Laboratories, 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/alexa fluor 568 goat anti rat igg intritrogen/product/Vector Laboratories
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    hrp conjugated goat anti mouse antibody  (Vector Laboratories)


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    Vector Laboratories hrp conjugated goat anti mouse antibody
    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 <t>peptide</t> <t>conjugated</t> RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using <t>streptavidin-HRP.</t> Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Hrp Conjugated Goat Anti Mouse Antibody, supplied by Vector Laboratories, 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/hrp conjugated goat anti mouse antibody/product/Vector Laboratories
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    hrp conjugated goat anti mouse antibody - by Bioz Stars, 2024-05
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    1) Product Images from "Surface-engineered extracellular vesicles for targeted delivery of therapeutic RNAs and peptides for cancer therapy"

    Article Title: Surface-engineered extracellular vesicles for targeted delivery of therapeutic RNAs and peptides for cancer therapy

    Journal: Theranostics

    doi: 10.7150/thno.68667

    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 peptide conjugated RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using streptavidin-HRP. Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Figure Legend Snippet: OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 peptide conjugated RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using streptavidin-HRP. Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.

    Techniques Used: Stable Transfection, Ligation, Functional Assay, Activity Assay, Western Blot, Competitive ELISA, Conjugation Assay, Incubation, Cryo-EM Sample Prep, Marker, One-tailed Test

    horseradish peroxidase hrp conjugated goat anti mouse igg antibody  (Vector Laboratories)


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    Vector Laboratories horseradish peroxidase hrp conjugated goat anti mouse igg antibody
    Horseradish Peroxidase Hrp Conjugated Goat Anti Mouse Igg Antibody, supplied by Vector Laboratories, 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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    peroxidase conjugated anti mouse  (Vector Laboratories)


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    Vector Laboratories peroxidase conjugated anti mouse
    Peroxidase Conjugated Anti Mouse, supplied by Vector Laboratories, 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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    horseradish peroxidase hrp conjugated affinity purified goat anti mouse igg h l  (Vector Laboratories)


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    Vector Laboratories horseradish peroxidase hrp conjugated affinity purified goat anti mouse igg h l
    Horseradish Peroxidase Hrp Conjugated Affinity Purified Goat Anti Mouse Igg H L, supplied by Vector Laboratories, 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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    hrp conjugated goat anti mouse igg  (Vector Laboratories)


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    Vector Laboratories hrp conjugated goat anti mouse igg
    Hrp Conjugated Goat Anti Mouse Igg, supplied by Vector Laboratories, 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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    hrp conjugated goat anti mouse igg  (Vector Laboratories)


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    Vector Laboratories hrp conjugated goat anti mouse igg
    Hrp Conjugated Goat Anti Mouse Igg, supplied by Vector Laboratories, 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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    secondary antibody hrp conjugated goat anti mouse igg was applied  (Vector Laboratories)


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    Vector Laboratories secondary antibody hrp conjugated goat anti mouse igg was applied
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    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 <t>peptide</t> <t>conjugated</t> RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using <t>streptavidin-HRP.</t> Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Hrp Conjugated Goat Anti Mouse Antibody, supplied by Vector Laboratories, 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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    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 <t>peptide</t> <t>conjugated</t> RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using <t>streptavidin-HRP.</t> Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Horseradish Peroxidase Hrp Conjugated Goat Anti Mouse Igg Antibody, supplied by Vector Laboratories, 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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    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 <t>peptide</t> <t>conjugated</t> RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using <t>streptavidin-HRP.</t> Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Horseradish Peroxidase Hrp Conjugated Affinity Purified Goat Anti Mouse Igg H L, supplied by Vector Laboratories, 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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    Vector Laboratories hrp conjugated goat anti mouse igg
    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 <t>peptide</t> <t>conjugated</t> RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using <t>streptavidin-HRP.</t> Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Hrp Conjugated Goat Anti Mouse Igg, supplied by Vector Laboratories, 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/hrp conjugated goat anti mouse igg/product/Vector Laboratories
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated goat anti mouse igg - by Bioz Stars, 2024-05
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    93
    Vector Laboratories secondary antibody hrp conjugated goat anti mouse igg was applied
    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 <t>peptide</t> <t>conjugated</t> RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using <t>streptavidin-HRP.</t> Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.
    Secondary Antibody Hrp Conjugated Goat Anti Mouse Igg Was Applied, supplied by Vector Laboratories, 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/secondary antibody hrp conjugated goat anti mouse igg was applied/product/Vector Laboratories
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    secondary antibody hrp conjugated goat anti mouse igg was applied - by Bioz Stars, 2024-05
    93/100 stars
      Buy from Supplier

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    OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 peptide conjugated RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using streptavidin-HRP. Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.

    Journal: Theranostics

    Article Title: Surface-engineered extracellular vesicles for targeted delivery of therapeutic RNAs and peptides for cancer therapy

    doi: 10.7150/thno.68667

    Figure Lengend Snippet: OaAEP1 ligase stably conjugates peptides onto the RBCEV surface. (A) Outline of the ligation reaction catalyzed by OaAEP1-Cys247Ala, where peptides with functional groups on the N-terminal are covalently ligated at the C-terminal to EV proteins. The inset provides a schematic representation of B-T140 peptide. The single letter code specifies amino acids responsible for targeting activity (green) or ligation (red). (B) Western blot (WB) analysis of B-T140 and B-TL5 peptides ligated to RBCEVs. (C) Copy number of B-TL5 and B-T140 peptides per EV determined using competitive ELISA (n = 3-6 biological replicates). (D) Percentage of peptide ligated EVs obtained via single-EV flow cytometric analysis of B-TL5 and B-T140 peptide conjugated RBCEVs, probed for biotin using streptavidin-AF488(n = 3 replicates). (E) Summary of western blot analysis assessing the stability of OaAEP1-mediated conjugation, based on the percentage of remaining peptide on RBCEVs that were ligated with B-TL5/B-T140 and incubated in EV-depleted human plasma at 37°C for 24-96 hrs (n = 3 replicates for each timepoint). (F) Irreversibility of OaAEP1 mediated ligation: B-T140 ligated RBCEVs were re-ligated with unbiotinylated T140 peptide repeatedly (repeat R1-3) and analyzed by western blot. The last lane illustrates EVs directly ligated with unbiotinylated T140 for reference. (G) Representative cryo-EM images of unligated and peptide-ligated RBCEVs. Scale bar, 100 nm. In B and F , the abundance of biotinylated peptide-conjugated proteins were quantified by the level of biotin detected using streptavidin-HRP. Molecular weights (kDa) of the protein marker are shown on the left of each blot. For quantification of peptides per EV in C , NTA was used to obtain the number of input EVs, thereby providing an estimate of peptide copy number per EV. The graphs present the mean ± SEM. Student's one-tailed t-test ***P < 0.001.

    Article Snippet: The wells were incubated for two hours with 100 μL HRP-conjugated goat anti-mouse antibody (Vector Bioscience) diluted at 1:3000 in blocking buffer.

    Techniques: Stable Transfection, Ligation, Functional Assay, Activity Assay, Western Blot, Competitive ELISA, Conjugation Assay, Incubation, Cryo-EM Sample Prep, Marker, One-tailed Test