Rsv f protein

Manufactured by Sino Biological

Sourced in United States

The RSV F protein is a recombinant protein produced in HEK293 cells. It is the fusion (F) protein of the respiratory syncytial virus (RSV), which is a key antigen for the development of RSV vaccines and diagnostics.

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Lab products found in correlation

Rsv g protein, by Sino Biological (3 mentions) Ag1478, by Merck Group (2 mentions) Neutralizing anti egfr ab 5 mab, by Merck Group (2 mentions) Ag 1295, by Santa Cruz Biotechnology (2 mentions) Interferon ifn lambda λ polyclonal abs, by Santa Cruz Biotechnology (2 mentions) Ifn λ receptor il 28r il 10rβ abs, by Santa Cruz Biotechnology (2 mentions) N propyl gallete, by Merck Group (2 mentions) Gefitinib, by Bio-Techne (2 mentions) Microtiter plate, by Greiner (1 mentions) Bicarbonate buffer, by Merck Group (1 mentions) Versamax plate reader, by Molecular Devices (1 mentions) 6 o desulfated heparin, by Iduron (1 mentions) Streptavidin sa sensor chips, by Cytiva (1 mentions) Biacore 3000, by Cytiva (1 mentions) 3 3 diaminobenzidine tetrahydrochloride substrate, by Thermo Fisher Scientific (1 mentions) Hrp conjugated anti mouse igg antibody, by Thermo Fisher Scientific (1 mentions) Ez read 400 microplate reader, by Harvard Bioscience (1 mentions) Capture elisa, by R&D Systems (1 mentions) O phenylenediamine, by Thermo Fisher Scientific (1 mentions) El800, by Agilent Technologies (1 mentions)

Spelling variants of this product

Recombinant RSV-F protein (3 citations)

7 protocols using rsv f protein

EGFR Inhibitor Impacts on Growth Factors

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Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (AG
1478), epidermal growth factor (EGF), and transforming growth factor
(TGF)-α, diphenyleneiodonium chloride (DPI), neutralizing anti-EGFR
(Ab-5) mAb, and an isotype-matched Ab were obtained from EMD Millipore
(Billerica, MA). Platelet-derived growth factor (PDGF) receptor tyrosine kinase
inhibitor (AG 1295), Janus kinase (Jak) 1 inhibitor, Interferon (IFN) lambda
(-λ) polyclonal Abs, IFN-λ receptor (IL-28R/IL-10Rβ) Abs,
and isotype-matched Abs were purchased from Santa Cruz Biotechnology (Santa
Cruz, CA). N-propyl gallete (nPG) was purchased from Sigma (St.
Louis, MO). Gefitinib was purchased from Tocris Biosciences (Bristol, UK). RSV F
protein was obtained from Sino Biological (Beijing, China).

Kalinowski A., Galen B.T., Ueki I.F., Sun Y., Mulenos A., Osafo-Addo A., Clark B., Joerns J., Liu W., Nadel J.A., Cruz C.S, & Koff J.L. (2018). Respiratory syncytial virus activates epidermal growth factor receptor to suppress interferon regulatory factor 1-dependent interferon-lambda and antiviral defense in airway epithelium. Mucosal immunology, 11(3), 958-967.

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Avidity Assay for RSV F Protein

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Avidity assay directed against F protein was performed by ELISA. Microtiter plates (Greiner Bio-one, cat. 655061) were coated with 1 µg/mL of RSV F protein (SinoBiologicals, cat. 11049-V08B) in bicarbonate buffer (Sigma, cat. C3041). Plates were incubated overnight at 4°C and then blocked with PBS-Tween 0.05%-milk 5% for 1 h.
Serum were serially diluted in blocking buffer and dispensed over three dilutions to frame an OD of 1 (based on F-specific IgG ELISA results) in eight replicates (one for each urea concentration). After 1.5 hour incubation at 37°C, plates were washed with PBS-Tween 0.05% and urea treatment (Sigma, cat. 51456) was performed with different concentrations of urea (0, 2, 3, 4, 5, 6, 7 and 8 M; 100 µL/well) for 10 min at RT and the reaction was stopped by washing the plates. Then, plates were incubated for 1.5 hours at 37°C with a goat anti-monkey IgG-HRP diluted 1:10,000 (Biorad, cat. AAI42P). Plates were washed and incubated with TMB substrate (Tebu-bio, cat. TMB100-1000) for 30 min in the dark at RT. Colorimetric reaction was stopped with 1 N HCl (VWR Prolabo, cat 30024290). OD were measured at 450–650 nm on a Versamax plate reader (Molecular Devices), the percentage of antibody binding was determined for each urea concentration as follows: OD[No Urea]/OD[Urea]*100 and AUC (Area under curves) was determined on Graph Pad Prism.

Pavot V., Bisceglia H., Guillaume F., Montano S., Zhang L., Boudet F, & Haensler J. (2021). A novel vaccine adjuvant based on straight polyacrylate potentiates vaccine-induced humoral and cellular immunity in cynomolgus macaques. Human Vaccines & Immunotherapeutics, 17(7), 2336-2348.

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Respiratory Syncytial Virus Glycoprotein Binding Kinetics

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RSV-G protein (Cat: 11070-V08H2), RSV-F protein (Cat: 40628-V08B) were purchased from Sino Biological Inc. The proteins were constructed as follows: 1) a DNA sequence encoding the glycoprotein G extracellular domain (Asn66-Arg297) of human respiratory syncytial virus A (93% homologous with strain rsb1734) (P27022-1) was expressed, with a C-terminal polyhistidine tag; and 2) a DNA sequence encoding the human respiratory syncytial virus fusion (AFX60213.1) (Met1-Ile525) was expressed with a polyhistidine tag at the C-terminus. Unfractionated heparin (15 kDa) was from Celsus Laboratories (Cincinnati, OH). Heparin oligosaccharides from tetrasaccharide (dp4) to octadecasaccharide (dp18) were from Iduron (Manchester, United Kingdom). Desulfated heparins including N-desulfated heparin (14 kDa), 2-O-desulfated IdoA heparin (13 kDa), 6-O-desulfated heparin (13 kDa) were from Iduron (Manchester, United Kingdom). Mucopolysaccharide polysulfate (MPS; 14.5 kDa) was from Luitpold Pharma (Munich, Germany). Pentosan polysulfate (PPS; 6.5 kDa) was from Bene Pharma (Munich, Germany). Sensor streptavidin (SA) chips were from Cytiva (Uppsala, Sweden). SPR experiments were performed using a BIAcore 3000 (Cytiva, Uppsala, Sweden) with Biaevaluation software (version 4.0.1).

Shi D., He P., Song Y., Linhardt R.J., Dordick J.S., Chi L, & Zhang F. (2023). Interactions of heparin with key glycoproteins of human respiratory syncytial virus. Frontiers in Molecular Biosciences, 10, 1151174.

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EGFR Inhibitor Impacts on Growth Factors

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Quantification of RSV-Specific Antibodies and Viral Loads

Cited 2 times

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RSV A2 virus-specific antibodies (IgG, IgG1, and IgG2a) were determined in sera by enzyme-linked immunosorbent assay (ELISA) using FI-RSV (4 μg/ml) [14 (link)], RSV F protein (100 ng/ml, BEI, NIAID, NIH), or RSV G protein (200 ng/ml, Sino biological Inc.) as a coating antigen. RSV-specific neutralizing antibody titers in mouse sera were measured using the red fluorescent monomeric Katushka 2 protein expressing RSV A2 strain (A2-K-line19F) as described [21 (link), 22 (link)]. Equal volumes of the diluted sera were mixed with A2-K-line19F virus to yield approximately 100 red-fluorescent (excitation at 588 nm, emission at 635 nm) RSV infected-cell spots per well. The percentages of red-fluorescent RSV infected-cells were presented as a measure of serum neutralization activity.
RSV titers in lung samples at day 5 post-challenge were determined using an immunoplaque assay as described [14 (link)]. Serially diluted lung homogenates were added to the HEp2 cell monolayer plates and incubated 3–6 days at 37°C. After fixing with ice-cold acetone-methanol and air drying, individual plaques were visualized and counted by anti-RSV F monoclonal antibody, HRP conjugated anti-mouse IgG antibody, and 3,3′-Diaminobenzidine tetrahydrochloride substrate (Invitrogen). The viral load detection limit is estimated to be approximately 60 PFU from each mouse lung sample in this assay.

Lee J.S., Kwon Y.M., Hwang H.S., Lee Y.N., Ko E.J., Yoo S.E., Kim M.C., Kim K.H., Cho M.K., Lee Y.T., Lee Y.R., Quan F.S, & Kang S.M. (2014). Baculovirus-expressed virus-like particle vaccine in combination with DNA encoding the fusion protein confers protection against respiratory syncytial virus. Vaccine, 32(44), 5866-5874.

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Murine IL-21 and Anti-RSV Antibody ELISA

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The concentrations of murine IL-21 were determined by capture ELISA (R&D Systems, USA), according to the manufacturer's instructions. Undetectable levels were considered zero for statistical purposes. For quantification of IgG and IgA RSV-specific antibodies in mice serum and bronchoalveolar lavage, 96-well plates were sensitized overnight with RSV F protein (Sino Biological Inc, USA), washed and blocked for 1 h with blocking buffer (5% milk + 0.05% tween in PBS 19 buffer) and serum or bronchoalveolar lavage was added in dilutions of 1/10, 1/100, 1/500, 1/1000, 1/ 10 000 and incubated for 2 h at room temperature. Rabbit antimouse IgG or IgA antibody, both horseradish peroxidase conjugated (Invitrogen, USA), followed by 3, 3', 5, 5' -Tetramethylbenzidine (TMB) (Life Technologies, Brazil) were used to develop the assay. Plates were read at 450 nm using an EZ Read 400 Microplate reader (Biochrom).
An ELISA to measure the avidity of anti-RSV antibody was conducted as described above; however, plates were washed with 6 M urea-PBS. The results were expressed as an Avidity Index, which was calculated as previously described. 6 The avidity of RSV-specific total IgG was classified according to values that had been predetermined arbitrarily, defined as low (<50%), intermediate (50-70%) or high (>70%).

Gassen R.B., Fazolo T., Nascimento de Freitas D., Borges T.J., Lima K., Antunes G.L., Maito F., Bueno Mendes D.A., Báfica A., Rodrigues LC J.r., Stein R., Duarte de Souza A.P, & Bonorino C. (2021). IL-21 treatment recovers follicular helper T cells and neutralizing antibody production in respiratory syncytial virus infection. Immunology and cell biology, 99(3).

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RSV Antibody Response Quantification

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RSV specific antibody responses were determined by enzyme-linked immunosorbent assay (ELISA) (Quan et al., 2011 (link)). Briefly, RSV-F protein (100 ng/ml, BEI, NIAID, NIH) or RSV-G protein (200 ng/ml, Sino biological) was used as an ELISA coating antigen. O-phenylenediamine (Invitrogen) was used as a substrate for horseradish peroxidase (HRP) conjugates of secondary antibodies. The optical density at 450 nm was read using an ELISA reader (BioTek, EL800).

Hwang H.S., Kwon Y.M., Lee J.S., Yoo S.E., Lee Y.N., Ko E.J., Kim M.C., Cho M.K., Lee Y.T., Jung Y.J., Lee J.Y., Li J.D, & Kang S.M. (2014). Co-immunization with virus-like particle and DNA vaccines induces protection against respiratory syncytial virus infection and bronchiolitis. Antiviral research, 110, 115-123.

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