Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Key resources and reagents

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Blocking Assay

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Key resources and reagents for the SB system

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques:

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Key resources and reagents for the ANG system

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques:

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Analyses of CA-p24 antigen concentration on diverse HIV isolates. Twenty-two HIV-1 isolates were tested through ABR-, ANG-, SB-, and RND-CA-p24 ELISAs for detection of CA-p24 antigen. All ELISAs detected most of the isolates, except RND-CA-p24 ELISA which did not detect the 92UG029 isolate. PBS was used as negative control. Values were measured in two independent ELISA runs and error bars represent the standard deviation (SD). The CA-p24 axis is log-scaled in the graph. The list of HIV isolates can be found in Table . ABR = Aalto Bio Reagents; ANG = Anogen; SB = Sino Biological; RND = R&D Systems

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Negative Control, Standard Deviation

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Fold-change in CA-p24 detection of HIV-1 A/B isolates relative to ABR-CA-p24 ELISA. ANG-CA-p24 ELISA exhibits low reactivity for HIV-1 A and B subtypes when compared to ABR-CA-p24 ELISA. RND-CA-p24 ELISA shows acceptable reactivity for HIV-1 A and B subtypes when compared to ABR-CA-p24 ELISA, but low reactivity for the SI22 isolate. SB-CA-p24 ELISA displays good reactivity for HIV-1 A and B subtypes when compared to ABR-CA-p24 ELISA, but low reactivity for the LAI isolate. ABR = Aalto Bio Reagents; ANG = Anogen; SB = Sino Biological; RND = R&D Systems

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Comparative reactivity towards HIV-1 isolates between assays as determined by Pearson’s r correlation. ANG-, SB-, and RND-CA-p24 ELISAs exhibit a significantly high correlation with ABR-CA-p24 ELISA. ANG-CA-p24 ELISA exhibits a significantly high correlation with RND-CA-p24 ELISA, whereas both RND- and ANG-CA-p24 ELISA show a low correlation with SB-CA-p24 ELISA. Concentrations of CA-p24 are displayed in ng/mL. ABR = Aalto Bio Reagents; ANG = Anogen; SB = Sino Biological; RND = R&D Systems

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Virologica Sinica

Article Title: Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

doi: 10.1016/j.virs.2023.06.005

Figure Lengend Snippet: Expression and S1/S2 cleavage efficiency of SARS-CoV-2 variants' spike proteins. A Expression of SARS-CoV-2 variants' spike proteins in 293T cells. mCherry fluorescence intensity indicates spike protein expression, and GFP fluorescence intensity is the reference control. The fluorescence intensity ratio of mCherry/GFP indicates the relative expression level. The Y axis was normalized with S-WT fluorescence intensity ratio value. The numbers labeled on the bars indicate the average fold change of variants' spike protein expression relative to the S-WT. Omicron indicates BA.1 sublineage (also termed B.1.1.529) here. B , C Spike protein cleavage of pseudovirus (PVs) of the WT and variants probed with antibody against RBD. HIV-1 p24 was used to normalize the protein sample loading. D , E S1/S2 cleavage efficiency of SARS-CoV-2 spikes. The spike proteins of WT and variants were expressed in HEK293T cells and at 48 ​h post-transfection, cell lysates were made for Western blot analysis using the antibody against RBD. GAPDH served as the loading control. Full-length spike (S0) and S1 proteins are indicated. The ratio of S1 to the total spike was quantitatively analyzed using ImageJ. Data represent the mean ​± ​SD of 3–5 independent replicates. Significance was analyzed by one-way ANOVA. P values less than 0.05 were considered to be statistically significant. ∗∗∗∗, P ​< ​0.0001; ∗∗∗, P ​< ​0.001; ∗∗, P ​< ​0.01; ∗, P ​< ​0.05; ns, not significant ( P ​> ​0.05).

Article Snippet: The following primary antibodies were used: mouse anti-SARS-CoV-2 spike protein antibody (Sino Biological, Cat: 40592-MM117), mouse anti-HIV-1 p24 protein monoclonal antibody (Sino Biological, Cat: 11695-MM15), and mouse anti-GAPDH antibody (Abcam, ab8245).

Techniques: Expressing, Fluorescence, Control, Labeling, Transfection, Western Blot

Journal: Virologica Sinica

Article Title: Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

doi: 10.1016/j.virs.2023.06.005

Figure Lengend Snippet: Cell entry ability of SARS-CoV-2 pseudotyped variants bearing mutate spike proteins. A The ability of SARS-CoV-2 pseudotyped variants entering into 293T-hACE2 cells (only expressing hACE2), 293T-hACE2-TMPRSS2 cells (co-expressing hACE2 and TMPRSS2), Caco-2 ​cells (co-expressing hACE2 and TMPRSS2) and 293T cells (expressing low hACE2). B Entry comparison of pseudotyped variants in cells overexpressing hACE2 and TMPRSS2 or without TMPRSS2. HIV-1 p24 was used to normalize the PVs' additive does, and the data was calculated at 5 ​ng p24. The numbers labeled on the bars indicate ( A ) the average fold change of PV variants relative to the PV-WT or ( B ) the average fold change of 293T-hACE2-TMPRSS2 cells relative to 293T-hACE2 cells. Data are mean ​± ​SD of 3 independent replicates. Mock, uninfected control (no virus); P values less than 0.05 were considered to be statistically significant. ∗∗∗∗, P ​< ​0.0001; ∗∗∗, P ​< ​0.001; ∗∗, P ​< ​0.01; ∗, P ​< ​0.05; ns, not significant ( P ​> ​0.05). Significance was analyzed by ( A ) one-way ANOVA or ( B ) t -test.

Article Snippet: The following primary antibodies were used: mouse anti-SARS-CoV-2 spike protein antibody (Sino Biological, Cat: 40592-MM117), mouse anti-HIV-1 p24 protein monoclonal antibody (Sino Biological, Cat: 11695-MM15), and mouse anti-GAPDH antibody (Abcam, ab8245).

Techniques: Expressing, Comparison, Labeling, Control, Virus

Journal: Virologica Sinica

Article Title: Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

doi: 10.1016/j.virs.2023.06.005

Figure Lengend Snippet: Cross-species infection of SARS-CoV-2 pseudotyped variants bearing mutate spike proteins. A The ability of SARS-CoV-2 pseudotyped variants entering into cells expressing ACE2 orthologs of human, mouse, mouse, rat, rhesus, Odocoileus virginianus and with TMPRSS2 (blue columns) or without (orange columns). B A heatmap shows the fold change of the cross-species infection performance of PV variants. The Control group is the ability of PV-WT to enter into 293T-hACE2 cells. The color bar represents the scale. HIV-1 p24 was used to normalize the PVs' additive does, and the data was calculated at 5 ​ng p24. Data are mean ​± ​SD of 3–4 replicates (four for Mu, and three for others) and were normalized to the WT of the individual experiment. Mock, uninfected control (no virus); P values less than 0.05 were considered to be statistically significant. ∗∗∗∗, P ​< ​0.0001; ∗∗∗, P ​< ​0.001; ∗∗, P ​< ​0.01; ∗, P ​< ​0.05; ns, not significant ( P ​> ​0.05). Significance was analyzed by one-way ANOVA.

Article Snippet: The following primary antibodies were used: mouse anti-SARS-CoV-2 spike protein antibody (Sino Biological, Cat: 40592-MM117), mouse anti-HIV-1 p24 protein monoclonal antibody (Sino Biological, Cat: 11695-MM15), and mouse anti-GAPDH antibody (Abcam, ab8245).

Techniques: Infection, Expressing, Control, Virus

Journal: Frontiers in Chemistry

Article Title: Identification of 6ω-cyclohexyl-2-(phenylamino carbonylmethylthio)pyrimidin-4(3 H )-ones targeting the ZIKV NS5 RNA dependent RNA polymerase

doi: 10.3389/fchem.2022.1010547

Figure Lengend Snippet: The interaction between 4w and ZIKV NS5 protein. (A) WB detection of ZIKV NS5 protein expression under different temperature gradients after 4w treatment; (B) NS5 protein grayscale analysis and ZIKV NS5 protein expression after 4w treatment observed changes in aggregation temperature; data is the mean (±SD) of three experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Subsequently, the protein was transferred to a polyvinylidene fluoride (PVDF) membrane and incubated with ZIKV E (1:2,000) and NS5 antibody (1:2,000) overnight at 4°C (Sino Biological, China) then add the corresponding secondary antibody and incubate at room temperature for 2 h. A specific signal was presented with a chemiluminescent substrate.

Techniques: Expressing

Journal: Frontiers in Chemistry

Article Title: Identification of 6ω-cyclohexyl-2-(phenylamino carbonylmethylthio)pyrimidin-4(3 H )-ones targeting the ZIKV NS5 RNA dependent RNA polymerase

doi: 10.3389/fchem.2022.1010547

Figure Lengend Snippet: Compound 4w inhibited the expression of ZIKV E and NS5 protein. (A) Western blot detected the inhibitory effect of 4w on ZIKV E and NS5 protein under the concentration gradient of 4w ; (B) Grayscale analysis and statistics of the inhibition of 4w on ZIKV E and NS5 protein under the concentration gradient of WB detection; Data is the mean (±SD) of three experiments, with DMSO as a positive contro * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. (C) Detected the inhibitory effect of 4w on ZIKV E protein by immunofluorescence.

Article Snippet: Subsequently, the protein was transferred to a polyvinylidene fluoride (PVDF) membrane and incubated with ZIKV E (1:2,000) and NS5 antibody (1:2,000) overnight at 4°C (Sino Biological, China) then add the corresponding secondary antibody and incubate at room temperature for 2 h. A specific signal was presented with a chemiluminescent substrate.

Techniques: Expressing, Western Blot, Concentration Assay, Inhibition, Immunofluorescence