rabbit anti gag p24 hiv 1 mab  (R&D Systems)

Journal: Virus Research

Article Title: Single amino acid substitution at position 614 in SARS-CoV-2 Spike Protein alters viral assembly and infectivity

doi: 10.1016/j.virusres.2025.199624

Figure Lengend Snippet: Effect of amino acid mutation at position 614 of S protein on viral packaging. (A and B) Western blot analysis of the protein expression of pseudovirus S protein and p24 in packaging cells and virus supernatant. (C) Schematic outline of the SARS-CoV-2 VLP production process. Plasmids encoding the SARS-CoV-2 structural proteins E, M, and N were cotransfected into HEK293T cells with plasmids encoding WT S or its mutants to obtain VLPs. (D and E) Western blot analysis of the expression of the major structural proteins S and NP on the VLPs. The original image is Supplemental Figure 4 in the supplementary material.

Article Snippet: GoldBand Plus 3-color Regular Range Protein Marker(8–180 kDa) (YEASEN, #20350ES72); SARS-CoV-2 (2019-nCoV) Spike RBD Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, # 40,592-T62); SARS-CoV-2 (2019-nCoV) Spike S2 Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, #40,590-T62); GFP (4B10) Mouse mAb (CST, #2955); Flag (DYKDDDDK) tag Polyclonal Antibody (Proteintech, #20,543–1-AP); Human Immunodeficiency Virus type 1 (HIV-1) Gag-p24 Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, #11,695-RP02); Myc-Tag (9B11) Mouse mAb (CST, #2276); GAPDH Mouse Monoclonal Antibody (Origene, #OTI2D9); β-actin Polyclonal Antibody (Proteintech, #20,536–1-AP).

Techniques: Mutagenesis, Western Blot, Expressing, Virus

Journal: Virus Research

Article Title: Single amino acid substitution at position 614 in SARS-CoV-2 Spike Protein alters viral assembly and infectivity

doi: 10.1016/j.virusres.2025.199624

Figure Lengend Snippet: Effect of amino acid mutation at position 614 of S protein on viral packaging. (A and B) Western blot analysis of the protein expression of pseudovirus S protein and p24 in packaging cells and virus supernatant. (C) Schematic outline of the SARS-CoV-2 VLP production process. Plasmids encoding the SARS-CoV-2 structural proteins E, M, and N were cotransfected into HEK293T cells with plasmids encoding WT S or its mutants to obtain VLPs. (D and E) Western blot analysis of the expression of the major structural proteins S and NP on the VLPs. The original image is Supplemental Figure 4 in the supplementary material.

Article Snippet: GoldBand Plus 3-color Regular Range Protein Marker(8–180 kDa) (YEASEN, #20350ES72); SARS-CoV-2 (2019-nCoV) Spike RBD Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, # 40,592-T62); SARS-CoV-2 (2019-nCoV) Spike S2 Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, #40,590-T62); GFP (4B10) Mouse mAb (CST, #2955); Flag (DYKDDDDK) tag Polyclonal Antibody (Proteintech, #20,543–1-AP); Human Immunodeficiency Virus type 1 (HIV-1) Gag-p24 Antibody, Rabbit PAb, Antigen Affinity Purified (Sinobiological, #11,695-RP02); Myc-Tag (9B11) Mouse mAb (CST, #2276); GAPDH Mouse Monoclonal Antibody (Origene, #OTI2D9); β-actin Polyclonal Antibody (Proteintech, #20,536–1-AP).

Techniques: Mutagenesis, Western Blot, Expressing, Virus

Journal: iScience

Article Title: Enhanced reverse zoonotic potential and immune evasion by omicron JN.1 variant

doi: 10.1016/j.isci.2025.112824

Figure Lengend Snippet: Transduction of various omicron S pseudovirions on 293/hACE2 and Calu3 cells (A) S proteins incorporation in pseudovirions. Pseudovirions with different omicron S proteins were pelleted down by centrifugation through 20% sucrose cushion and separated in a 10% SDS-PAGE. Detection of S proteins in pseudovirions was performed by Western blot using rabbit polyclonal anti-S2 antibodies. The p24 served as the loading controls. Experiments were done three times and one representative was shown. (B and C) Entry of pseudovirions of omicron variants. HEK 293/hACE2 and Calu3 cells were transduced with omicron S pseudovirions and lysed at 40 h post-transduction. The transduction efficiencies were determined according to luciferase activities. Experiments were done three times in triplicate and one representative is shown. The statistical difference relative to WT was determined by one-way ANOVA with Dunnett’s multiple testing correction ( n = 3). The statistical difference between BA.2.86 and JN.1 was determined by a two-tailed t-test ( n = 3). Error bars indicate SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Rabbit polyclonal against SARS-CoV-2 S2 antibodies (Cat#40509-T62), rabbit polyclonal against HIV-1 Gag-p24 antibodies (Cat#11695-T62), mouse polyclonal against FLAG tag antibodies (Cat#109143-MM13) and mouse polyclonal against beta-actin antibodies (Cat#100166-MM10) for western blotting and chimeric monoclonal against SARS-CoV-2 S2 antibody (Cat#40590-D001) used for flow cytometry was purchased from SinoBiological Inc (Beijing, China).

Techniques: Transduction, Centrifugation, SDS Page, Western Blot, Luciferase, Two Tailed Test

Journal: iScience

Article Title: Enhanced reverse zoonotic potential and immune evasion by omicron JN.1 variant

doi: 10.1016/j.isci.2025.112824

Figure Lengend Snippet: Transduction of various omicron S pseudovirions on 293/hACE2 and Calu3 cells (A) S proteins incorporation in pseudovirions. Pseudovirions with different omicron S proteins were pelleted down by centrifugation through 20% sucrose cushion and separated in a 10% SDS-PAGE. Detection of S proteins in pseudovirions was performed by Western blot using rabbit polyclonal anti-S2 antibodies. The p24 served as the loading controls. Experiments were done three times and one representative was shown. (B and C) Entry of pseudovirions of omicron variants. HEK 293/hACE2 and Calu3 cells were transduced with omicron S pseudovirions and lysed at 40 h post-transduction. The transduction efficiencies were determined according to luciferase activities. Experiments were done three times in triplicate and one representative is shown. The statistical difference relative to WT was determined by one-way ANOVA with Dunnett’s multiple testing correction ( n = 3). The statistical difference between BA.2.86 and JN.1 was determined by a two-tailed t-test ( n = 3). Error bars indicate SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Rabbit polyclonal against HIV-1 Gag-p24 antibodies , SinoBiological Inc (Beijing, China) , Cat#11695-T62.

Techniques: Transduction, Centrifugation, SDS Page, Western Blot, Luciferase, Two Tailed Test

Journal: bioRxiv

Article Title: HIV-1 Envelope glycoprotein modulates CXCR4 clustering and dynamics on the T cell membrane

doi: 10.1101/2025.06.11.659074

Figure Lengend Snippet: A) Representative images of clarified VLPs visualized by STED microscopy. Upper panels show images of the indicated VLPs stained for Gag p24 (blue) and gp120 (red). Lower panels show 10× magnification of equivalent images. White arrows indicate mature VLPs (p24 condensation). ( B) Percentage of mature VLPs, analyzed from the images in A) using TrackAnalyzer in ImageJ, based on p24 intensity and aggregation level (mean ± SD; n = 2; ****p ≤ 0.0001; the significance indicated on immature VLPs bar shows the difference with all other conditions). C) Percentage of VLPs expressing gp120 on their surface, as analyzed in ImageJ (mean ± SD; n = 2; ***p ≤ 0.001). D) Distribution of gp120 mean fluorescence intensity. Each spot corresponds to the mean fluorescence intensity for each analyzed VLP in a.u. The black line represents the mean of all values (**** p ≤ 0.0001). E) Frequency of gp120 intensity/particle. Statistical significance was determined by one-way-ANOVA followed by Tukey’s multiple comparisons test in panels B and C and by Mann-Whitney analysis for panel D.

Article Snippet: The following antibodies were used: anti-human CXCR4 monoclonal antibody (mAb) (clone 44717) and phycoerythrin-conjugated anti-human CXCR4 mAb (clone 12G5; both from R&D Systems, Minneapolis, MN); goat F(ab’)2 anti-mouse IgG-PE (Southern Biotech, Birmingham, AL); anti-human CD4 mAb (clone OKT4; Biolegend, San Diego, CA); anti-histidine mAb (clone AD1.1.10; R&D Systems); rabbit anti-gp120 IIIb Ab ; rabbit anti-Gag p24 HIV-1 mAb (R&D Systems); and anti-phospho-AKT mAb (S473; #4060), anti-phospho-ERK1,2 mAb (T202/Y204; #9191) and anti-phospho-Lck mAb (Y505; #2751) (all from Cell Signaling Technology, Danvers, MA); anti-tubulin mAb conjugated with rhodamine (Bio-Rad, Hercules, CA); phalloidin-TRITC (#P1951, Sigma-Merck, St Louis, MO); anti-ICAM 3 mAb (clone HP2/19) kindly donated by Dr. Francisco Sánchez Madrid; goat anti-mouse-AF488 Ab (Thermo Fisher Scientific); anti-human gp120 mAb Fab fragments (clone 2G12; Polymun Scientific, Vienna, Austria); anti-human IgG Fab fragments (Jackson ImmunoResearch, West Grove, PA) conjugated to Abberior STAR RED (Abberior GmbH, Gottingen, Germany) kindly donated by Dr. Jakub Chojnacki; anti-p24 HIV-1 (clone 37G12; Polymun Scientific, Vienna, Austria) conjugated with Abberior STAR ORANGE.

Techniques: Microscopy, Staining, Expressing, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: HIV-1 Envelope glycoprotein modulates CXCR4 clustering and dynamics on the T cell membrane

doi: 10.1101/2025.06.11.659074

Figure Lengend Snippet: The presence of CXCR4 R334X on JKCD4 + cells does not alter gp120 binding and increases fusion events with target cells expressing HIV pHXB2 envelope. A) Binding of X4-gp120 to target cells expressing CD4 and CXCR4 or CD4 and CXCR4 R334X analyzed by flow cytometry. Cells were incubated with 0.3 μg/mL of X4-gp120 at 37°C for 30 minutes. Data show MFI (arbitrary units, a.u.) mean ± SD; (n = 2). Statistical significance was determined using Student’s t-test (n.s.= not significant). B) Cell-cell fusion between JKHXBc2-expressing HIV-1 envelope and different target cells (JKCD4 + CXCR4 + , JKCD4 + CXCR4 − and JKCD4 + CXCR4 R334X ). Prior to co-culture, each cell type was loaded with the corresponding cell-tracker. Data show the percentage of fusion events ± SD (n = 6). We used as reference the fusions events detected in JKCD4 + CXCR4 + cells (100%). Statistical significance was determined by one-way-ANOVA (*p < 0.05, ****p ≤ 0.0001). C ) Representative biparametric histograms from cells in B showing CMAC versus orange fluorophores. D ) Human PBMCs isolated from a WHIM patient (WHIM) and three healthy donors (HD1-3) in two independent experiments were infected with X4-pseudotyped HIV-1 NL4-3 (MOI: 0.001). At 2 hours post infection (p.i.), supernatant samples were obtained at different time points (days post-infection) and p24 levels (pg/mL) in each sample were determined using a commercial ELISA. Results show mean ± SD (n = 2).

Article Snippet: The following antibodies were used: anti-human CXCR4 monoclonal antibody (mAb) (clone 44717) and phycoerythrin-conjugated anti-human CXCR4 mAb (clone 12G5; both from R&D Systems, Minneapolis, MN); goat F(ab’)2 anti-mouse IgG-PE (Southern Biotech, Birmingham, AL); anti-human CD4 mAb (clone OKT4; Biolegend, San Diego, CA); anti-histidine mAb (clone AD1.1.10; R&D Systems); rabbit anti-gp120 IIIb Ab ; rabbit anti-Gag p24 HIV-1 mAb (R&D Systems); and anti-phospho-AKT mAb (S473; #4060), anti-phospho-ERK1,2 mAb (T202/Y204; #9191) and anti-phospho-Lck mAb (Y505; #2751) (all from Cell Signaling Technology, Danvers, MA); anti-tubulin mAb conjugated with rhodamine (Bio-Rad, Hercules, CA); phalloidin-TRITC (#P1951, Sigma-Merck, St Louis, MO); anti-ICAM 3 mAb (clone HP2/19) kindly donated by Dr. Francisco Sánchez Madrid; goat anti-mouse-AF488 Ab (Thermo Fisher Scientific); anti-human gp120 mAb Fab fragments (clone 2G12; Polymun Scientific, Vienna, Austria); anti-human IgG Fab fragments (Jackson ImmunoResearch, West Grove, PA) conjugated to Abberior STAR RED (Abberior GmbH, Gottingen, Germany) kindly donated by Dr. Jakub Chojnacki; anti-p24 HIV-1 (clone 37G12; Polymun Scientific, Vienna, Austria) conjugated with Abberior STAR ORANGE.

Techniques: Binding Assay, Expressing, Flow Cytometry, Incubation, Co-Culture Assay, Isolation, Infection, Enzyme-linked Immunosorbent Assay

Journal: Cell Discovery

Article Title: A potent and broad CD4 binding site neutralizing antibody with strong ADCC activity from a Chinese HIV-1 elite neutralizer

doi: 10.1038/s41421-025-00808-x

Figure Lengend Snippet: a , b Neutralization potency ( a ) and breadth ( b ) of FD22, in comparison with VRC01, against a 145-isolate Env-pseudovirus panel. c , d The neutralization activity ( c ) and percentage ( d ) of FD22 were evaluated against different clades of HIV-1 pseudoviruses from a 145-virus panel, including the two predominant circulating strains in China, CRF01_AE and CRF07_BC. VRC01 served as a control. e Activation of HIV-1 latency in ACH-2 cells. HIV-1 latency reversal in ACH-2 cells was induced by treatment with TNF-α or panobinostat for 48 h. Viral reactivation was assessed by quantifying p24 antigen concentrations using a standardized ELISA. f Inhibitory activity of FD22 against latent HIV-1 in ACH-2 cells activated by TNF-α or panobinostat, as measured in TZM-bl cells. ACH-2 cells were treated with TNF-α (10 ng/mL) or panobinostat (1 μM) to induce HIV-1 reactivation. After activation, FD22 was added at various concentrations to assess its inhibitory activity. Reactivated virus production was measured by coculturing supernatants from treated ACH-2 cells with TZM-bl indicator cells.

Article Snippet: Rabbit polyclonal anti-Gag-p24 antibodies (Sino Biological Inc.) were used to coat ELISA plates.

Techniques: Neutralization, Comparison, Activity Assay, Virus, Control, Activation Assay, Enzyme-linked Immunosorbent Assay