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Anti vsv g

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Anti-VSV-G is a lab equipment product. It functions as an antibody that specifically binds to the Vesicular Stomatitis Virus Glycoprotein (VSV-G), a protein found on the surface of the Vesicular Stomatitis Virus.

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Lab products found in correlation

Sars cov 2 b 1, by Addgene (6 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) Anti flag, by Merck Group (3 mentions) Anti ha clone ha 7, by Merck Group (3 mentions) Protran, by Cytiva (3 mentions) Vsv g, by Addgene (2 mentions) Sars cov 2 wuhan hu 1, by Addgene (2 mentions) Sars cov 2 p 1, by Addgene (2 mentions) Sars cov 1, by Addgene (2 mentions) Mers cov, by Addgene (2 mentions) Rvsvδg g gfp virus, by Kerafast (2 mentions) Anti flag m2, by Merck Group (2 mentions) Anti ha, by Merck Group (2 mentions) Mini protean system, by Bio-Rad (2 mentions) Mouse secondary antibodies, by Merck Group (2 mentions) Anti flag clone m2, by Merck Group (2 mentions) Anti strepii, by Merck Group (2 mentions) Anti 5his, by Merck Group (2 mentions) Rolipram, by Merck Group (1 mentions) Isoproterenol hydrochloride, by Merck Group (1 mentions)

Spelling variants of this product

VSV-G (10 citations) Rabbit anti-VSV-G (8 citations) Anti-VSV-G antibody (4 citations) Anti‐VSV (3 citations) Anti-VSV-G Agarose Conjugate (2 citations) Anti‐VSV‐G (clone P5D4 (2 citations)

16 protocols using anti vsv g

1

Production of VSV-dG Pseudoviral Particles

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VSV-dG pseudoviral particles were produced as previously described [34 (link)]. Briefly, 8 × 106 HEK293T cells were plated in 10-cm tissue culture dishes and transfected using Lipofectamine2000 (Invitrogen) with plasmids encoding different CoV spike proteins or VSV-G protein. Expression vectors for SARS-CoV-2 Wuhan-Hu-1 (Addgene, #149539), SARS-CoV-2 B.1.167.2 (Addgene, #172320), SARS-CoV-2 B.1.1.7 (Addgene, #170451), SARS-CoV-2 B.1.351 (Addgene, #170449), SARS-CoV-2 P.1 (Addgene, #170450), SARS-CoV-1 (Addgene, #170447), MERS-CoV (Addgene, #170448), and VSV-G (Addgene, #12259) were used. At 24 h posttransfection, cells were incubated with replication restricted rVSVΔG*G-GFP virus (Kerafast, #EH1019-PM) at approximately 5 MOI for 1 h at 37°C, 5% CO2 and the media was replaced with complete media. Anti-VSV-G (Sigma, #MABF2337) was added at final concentration of 1 μg/mL to neutralize residual rVSVΔG*G. At approximately 24 h postinoculation, viral supernatant was harvested, cell debris were removed by centrifuging for 10 min at 1,320 × g, and stored at −80°C in small aliquots.
Song J., Chow R.D., Peña-Hernández M.A., Zhang L., Loeb S.A., So E.Y., Liang O.D., Ren P., Chen S., Wilen C.B, & Lee S. (2022). LRRC15 inhibits SARS-CoV-2 cellular entry in trans. PLOS Biology, 20(10), e3001805.
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2

Characterization of cAMP Signaling Modulators

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GIBCO cell culture media, serum, and antibiotics were obtained from Invitrogen. All chemicals were obtained from Sigma: a general PDE inhibitor IBMX (Cat# I5879); a specific PDE4 inhibitor rolipram (Cat# R6520); a specific PDE3 inhibitor cilostamide (Cat# C7971); β-AR agonist isoproterenol hydrochloride (ISO, Cat# I6504); PKA inhibitor fragment 14–22, myristoylated trifluoroacetate salt (PKI, Cat# P9115); forskolin (FSK, F6886); H89 dihydrochloride hydrate (H89, B1427), vehicle control dimethyl sulfoxide (DMSO, Cat# D4540). Antibodies used were as follows: anti-α-actinin (clone BM-75.2, Sigma, Cat# A5044), anti-Flag (M2, Sigma, Cat# F1804), anti-VSV-G (Sigma, Cat# V4888,), anti-V5-agarose affinity gel (V5-10, Sigma, Cat# A7345), anti-Myc (4A6, EMD Millipore, Cat# 05-724), anti-V5 (Invitrogen, Cat# R96025), anti-GFP (Clontech, Cat# 632375 and Cat# 632592), anti-phospho-CREB at Ser133 (87G3, cell signaling, Cat# 9198), anti-CREB (48H2, cell signaling, Cat#3955). AKAP-Lbc polyclonal antibody VO96 were previously described [44 (link)].
Wang L., Burmeister B.T., Johnson K.R., Baillie G.S., Karginov A.V., Skidgel R.A., O’Bryan J.P, & Carnegie G.K. (2015). UCR1C is a novel activator of phosphodiesterase 4 (PDE4) long isoforms and attenuates cardiomyocyte hypertrophy. Cellular signalling, 27(5), 908-922.
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3

Western Blot Protein Detection Protocol

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Cell lysates were separated by SDS-PAGE on 4–12% or 12% Bis-Tris NuPAGE gels in MES or MOPS running buffer (Life Technologies). Either the iBlot dry blotting system or the XCell II Blot Module (Life Technologies) was used to transfer proteins to either PVDF or nitrocellulose. Membranes were blocked with SuperBlock Blocking Buffer (Pierce) with 0.25% Surfact-Amps 20 (Pierce) for 1 hour to overnight. Membranes were then probed with polyclonal anti-VSV-G (diluted 1:1,500; Sigma) or anti-GroEL (diluted 1:160,000; provided by Karsten Hazlett, Albany Medical College, Albany, New York, United States) for one hour, washed (10 minutes incubations in TBST plus 0.25% Surfact-Amps 20, 4 times) and re-blocked for 1 hour. After membranes were incubated with polyclonal goat anti-rabbit (diluted 1:10,000; Pierce) and washed, proteins were detected using SuperSignal West Pico Chemiluminescent Substrate (Life Technologies).
Ramsey K.M., Osborne M.L., Vvedenskaya I.O., Su C., Nickels B.E, & Dove S.L. (2015). Ubiquitous Promoter-Localization of Essential Virulence Regulators in Francisella tularensis. PLoS Pathogens, 11(4), e1004793.
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4

Western Blot Analysis of Plexin Proteins

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shRNAs and cDNA constructs were transiently transfected into HEK293T cells, and after 48–72hrs these cells were harvested in 3x loading buffer + 10% β-mercaptoethanol. Lysates from cells transfected with Plexin-B1 and Plexin-B2 cDNAs were loaded onto a 6% SDS-PAGE gel for electrophoresis, and all other Westerns used a 10% SDS-PAGE gel. Proteins were transferred to a nitrocellulose membrane at 4°C overnight, blocked in 5% milk/TBST solution for 1 hr, then incubated overnight at 4˚C in primary antibody [anti- VSV-g (1:100; Sigma, V4888); anti- myc (Santa Cruz; sc-40; 1:500)]. The following day the membrane was washed three times for 15 min in TBST and then incubated with secondary antibody [anti- rabbit IR680LT (Licor; 1:10,000); anti-mouse IR800CW (Licor; 1:10,000)] for 2 hr at room temperature. After three 15 minute washes in TBST, fluorescent images were obtained using the Odyssey Western Blotting System (Licor). To assess loading, blots were probed with anti-mouse Tubulin (1:5000; Sigma, T5168) and the secondary antibody anti-mouse IR800CW (1:10,000; Licor) similar to above.
McDermott J.E., Goldblatt D, & Paradis S. (2018). Class 4 Semaphorins and Plexin-B receptors regulate GABAergic and glutamatergic synapse development in the mammalian hippocampus. Molecular and cellular neurosciences, 92, 50-66.
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5

Immunoblotting of Bacterial Proteins

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Proteins suspended in loading buffer were subjected to SDS-PAGE. For detection by immunostaining, proteins were transferred onto nitrocellulose membranes, and immunoblots were probed with primary antibodies, and goat secondary antibodies coupled to alkaline phosphatase, and developed in alkaline buffer in presence of 5-bromo-4-chloro-3-indolylphosphate and nitroblue tetrazolium. Anti-TolA,-TolB,-Pal and-OmpA polyclonal antibodies are from our laboratory collection. Anti-FLAG (Sigma-Aldrich), anti-VSV-G (Sigma-Aldrich), anti-HA (Roche), anti-EFTu (Hycult Biotech) and anti-rabbit,-mouse or-rat alkaline phosphatase-conjugated goat secondary antibodies (Beckman Coulter) have been purchased as indicated and used as recommended by the manufacturer.
Brunet Y.R., Zoued A., Boyer F., Douzi B, & Cascales E. (2015). The Type VI Secretion TssEFGK-VgrG Phage-Like Baseplate Is Recruited to the TssJLM Membrane Complex via Multiple Contacts and Serves As Assembly Platform for Tail Tube/Sheath Polymerization. PLoS Genetics, 11(10), e1005545.
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6

Epitope-tagged Cell Line Immunoprecipitation

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Immunoprecipitation began with 1 to 3 confluent 13-ml tubes per epitope-tagged cell line. After detachment by icing, cells were pelleted at 700 × g and washed once in HBS. The cells were resuspended in 300 μl of lysis buffer (50 mM Tris [pH 7.5], 150 mM NaCl, 7.5% glycerol, 0.25 mM CaCl2, 0.25 mM ATP, 0.05 mM DTT, 0.5 mM PMSF, 0.1% Triton X-100, 2× HALT protease inhibitors [Pierce]) and sonicated. The lysate was cleared by centrifugation at 10,000 × g for 10 min at 4°C and added to 30 μl of lysis buffer-equilibrated anti-HA (Sigma), anti-VSVG (Sigma), or StrepTactin (IBA) beads. After 1.5 h of binding, the beads were washed four times with wash buffer (25 mM Tris [pH 7.5], 150 mM NaCl, 0.25 mM CaCl2, 0.25 mM ATP, 5% glycerol, 0.05% Tween 20) and then boiled in 50 μl of sample buffer. Immunoblotting was performed as described below.
Krtková J., Xu J., Lalle M., Steele-Ogus M., Alas G.C., Sept D, & Paredez A.R. (2017). 14-3-3 Regulates Actin Filament Formation in the Deep-Branching Eukaryote Giardia lamblia. mSphere, 2(5), e00248-17.
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7

Immunoblotting Techniques Using Various Antibodies

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SDS–PAGE was performed on Bio-Rad Mini-PROTEAN systems using standard protocols. For immunostaining, proteins were transferred onto 0.2-μm nitrocellulose membranes (Amersham Protran) Immunoblots were probed with primary antibodies and goat secondary antibodies coupled to alkaline phosphatase and developed in alkaline buffer in presence of 5-bromo-4-chloro-3- indolylphosphate and nitro-blue tetrazolium. The anti-HA (HA-7 clone, Sigma Aldrich), anti-Flag (Sigma Aldrich, A2220-1ml or F3165-.2MG), anti-StrepII (Bio-Rad), anti VSV-G (Sigma Aldrich, SAB4200695-100UL), anti-6His (Sigma Aldrich, SAB1305538-400UL), anti-StrepII (Biorad MCA2489), anti-GFP (Thermofischer, MA5-15256), RNA-pol II (Ozyme 664906) monoclonal antibodies, rabbit polyclonal mcherry antibodies (OriGene TA150125) mouse secondary antibodies (Interchim 115-055-003) and rat secondary antibodies (Interchim115-055-045) were purchased as indicated. Synthetics polyclonalrabbit antibodies were designed by GeneScript for Hcp (epitope: SVGGHTAERVEHSDC) and TssM detection (epitope: CAPAQAAAPAKTENP).
Kandolo O., Cherrak Y., Filella-Merce I., Le Guenno H., Kosta A., Espinosa L., Santucci P., Verthuy C., Lebrun R., Nilges M., Pellarin R, & Durand E. (2023). Acinetobacter type VI secretion system comprises a non-canonical membrane complex. PLOS Pathogens, 19(9), e1011687.
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8

Production of Pseudoviral Particles Expressing Different SARS-CoV-2 Spike Proteins

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VSV-dG pseudoviral particles were produced as previously described [31 (link)]. Briefly, 8 × 106 HEK293T cells were plated in 10-cm tissue culture dishes and transfected using Lipofectamine2000 (Invitrogen) with plasmids encoding different CoV spike proteins or VSV-G protein. Expression vectors for SARS-CoV-2 Wuhan-Hu-1 (Addgene, #149539), SARS-CoV-2 B.1.167.2 (Addgene, #172320), SARS-CoV-2 B.1.1.7 (Addgene, #170451), SARS-CoV-2 B.1.351 (Addgene, #170449), SARS-CoV-2 P.1 (Addgene, #170450), SARS-CoV-1 (Addgene, #170447), MERS-CoV (Addgene, #170448) and VSV-G (Addgene, #12259) were used. At 24 h post transfection, cells were incubated with replication restricted rVSVΔG*G-GFP virus (Kerafast, #EH1019-PM) at ~5 MOI for 1 h at 37°C, 5% CO2 and the media was replaced with complete media. Anti-VSV-G (Sigma, #MABF2337) was added at final concentration of 1 μg/mL to neutralize residual rVSVΔG*G. At ~24 h post inoculation, viral supernatant was harvested, cell debris were removed by centrifuging for 10 min at 1320 × g, and stored at −80°C in small aliquots.
Song J., Chow R.D., Pena-Hernandez M., Zhang L., Loeb S.A., So E.Y., Liang O.D., Wilen C.B, & Lee S. (2021). LRRC15 is an inhibitory receptor blocking SARS-CoV-2 spike-mediated entry in trans. bioRxiv.
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9

Investigating Innate Immune Signaling Pathways

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Human embryonic kidney cells (HEK293T) and A549 cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Invitrogen) supplemented with 10% FBS, 10 mM Hepes and 2 mM L-glutamine. Poly(I:C) and Poly(dA:dT) were purchased from InvivoGen. The silver staining kit and Lipofectamine 2000 were purchased from Invitrogen. The following antibodies were used for immunoprecipitation and/or immunoblotting: anti-DDX23 (abcam), anti-TRIF (Cell signaling technology, CST), MAVS (abcam), TBK1, p-TBK1, p38, p-p38, p65, p-p65, IRF3, p-IRF3 (CST). Anti-HA, anti-FLAG beads, Poly(I:C) agarose, poly(C) agarose, and anti-VSV-G were purchased from Sigma.
Ruan J., Cao Y., Ling T., Li P., Wu S., Peng D., Wang Y., Jia X., Chen S., Xu A, & Yuan S. (2019). DDX23, an Evolutionary Conserved dsRNA Sensor, Participates in Innate Antiviral Responses by Pairing With TRIF or MAVS. Frontiers in Immunology, 10, 2202.
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10

Protein Separation and Detection

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Proteins were separated on sodiumdodecyl sulfate (SDS)-polyacrylamide gels (10% acrylamide) and either visualised by silver staining using the Silver Stain PlusOne kit (GE Healthcare, Little Chalfont, UK) or SilverXpress kit (Invitrogen) or transferred to nitrocellulose membranes (Hybond enhanced chemiluminescence, ECL, Amersham). Antibodies used were: anti-p24 (1:100, mouse monoclonal, ARP 365, CFAR, NIBSC, UK; binds sequence NPPIPVGEIY in p24 of HIV-1 Gag), anti-VSV-G (1:2000, mouse monoclonal, Sigma-Aldrich), anti-RDpro pg70 (1:2000, goat polyclonal, Quality Biotech Inc., Camden, NJ, USA), anti-AHANAK (1:500, mouse-monoclonal, Abcam, Cambridge, UK), anti-TSG101 (1:500, rabbit-polyclonal, Sigma-Aldrich), anti-ALIX (1:1000, rabbit polyclonal), anti-EEF1A (1:500, mouse-monoclonal), anti-ENO1 (1:500, mouse-monoclonal), anti-MARCKSL1 (1:1000, rabbit-polyclonal) and anti-GAPDH (1:5000, mouse-monoclonal, all by Millipore, Billerica, MA, USA).
Johnson S., Wheeler J.X., Thorpe R., Collins M., Takeuchi Y, & Zhao Y. (2018). Mass spectrometry analysis reveals differences in the host cell protein species found in pseudotyped lentiviral vectors. Biologicals, 52, 59-66.
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