chikv antigen  (ATCC)


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  • 93
    Name:
    Chikungunya virus immune ascitic fluid V 548 701 562
    Description:
    Source Mouse
    Catalog Number:
    VR-1241AF
    Price:
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    Source:
    Mouse
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    Structured Review

    ATCC chikv antigen
    Source Mouse
    https://www.bioz.com/result/chikv antigen/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    chikv antigen - by Bioz Stars, 2021-07
    93/100 stars

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    Related Articles

    High Performance Liquid Chromatography:

    Article Title: Chikungunya Virus Strains from Each Genetic Clade Bind Sulfated Glycosaminoglycans as Attachment Factors
    Article Snippet: Slides were blocked at room temperature for 1 h with HBS buffer (10 mM HEPES at pH 7.4 with 150 mM NaCl and 5 mM CaCl2) supplemented to contain 0.02% (wt/vol) casein (Pierce) and 1% (wt/vol) bovine serum albumin (BSA) (Sigma). .. Microarrays were overlaid with VLP solution (50 μg/ml was used in most analyses) at room temperature for 1.5 h and fixed with 4% paraformaldehyde (PFA) diluted in high-pressure liquid chromatography (HPLC)-grade water at 4°C for 30 min. VLP binding was detected following incubation with anti-CHIKV E2 antibody (CHK-152 [ ]; 1:300) or ascites fluid (ATCC VR-1241AF; 1:300) at room temperature for 1 h, biotinylated goat anti-mouse IgG (Sigma; 2 μg/ml) at room temperature for 1 h, and Alexa Fluor 647-labeled streptavidin (Molecular Probes;1 μg/ml) at room temperature for 30 min. .. Imaging and data analysis are described in the supplementary MIRAGE document (Table S3).

    Binding Assay:

    Article Title: Chikungunya Virus Strains from Each Genetic Clade Bind Sulfated Glycosaminoglycans as Attachment Factors
    Article Snippet: Slides were blocked at room temperature for 1 h with HBS buffer (10 mM HEPES at pH 7.4 with 150 mM NaCl and 5 mM CaCl2) supplemented to contain 0.02% (wt/vol) casein (Pierce) and 1% (wt/vol) bovine serum albumin (BSA) (Sigma). .. Microarrays were overlaid with VLP solution (50 μg/ml was used in most analyses) at room temperature for 1.5 h and fixed with 4% paraformaldehyde (PFA) diluted in high-pressure liquid chromatography (HPLC)-grade water at 4°C for 30 min. VLP binding was detected following incubation with anti-CHIKV E2 antibody (CHK-152 [ ]; 1:300) or ascites fluid (ATCC VR-1241AF; 1:300) at room temperature for 1 h, biotinylated goat anti-mouse IgG (Sigma; 2 μg/ml) at room temperature for 1 h, and Alexa Fluor 647-labeled streptavidin (Molecular Probes;1 μg/ml) at room temperature for 30 min. .. Imaging and data analysis are described in the supplementary MIRAGE document (Table S3).

    Incubation:

    Article Title: Chikungunya Virus Strains from Each Genetic Clade Bind Sulfated Glycosaminoglycans as Attachment Factors
    Article Snippet: Slides were blocked at room temperature for 1 h with HBS buffer (10 mM HEPES at pH 7.4 with 150 mM NaCl and 5 mM CaCl2) supplemented to contain 0.02% (wt/vol) casein (Pierce) and 1% (wt/vol) bovine serum albumin (BSA) (Sigma). .. Microarrays were overlaid with VLP solution (50 μg/ml was used in most analyses) at room temperature for 1.5 h and fixed with 4% paraformaldehyde (PFA) diluted in high-pressure liquid chromatography (HPLC)-grade water at 4°C for 30 min. VLP binding was detected following incubation with anti-CHIKV E2 antibody (CHK-152 [ ]; 1:300) or ascites fluid (ATCC VR-1241AF; 1:300) at room temperature for 1 h, biotinylated goat anti-mouse IgG (Sigma; 2 μg/ml) at room temperature for 1 h, and Alexa Fluor 647-labeled streptavidin (Molecular Probes;1 μg/ml) at room temperature for 30 min. .. Imaging and data analysis are described in the supplementary MIRAGE document (Table S3).

    Article Title: A Single-Amino-Acid Polymorphism in Chikungunya Virus E2 Glycoprotein Influences Glycosaminoglycan Utilization
    Article Snippet: Infected cells were visualized using indirect immunofluorescence. .. Cells were incubated in blocking buffer (phosphate-buffered saline [PBS; Gibco] containing 5% FBS and 0.1% Triton X-100) at room temperature for 1 h and stained with precleared anti-CHIKV immune ascetic fluid (ATCC VR-1241AF) diluted 1:1,500 and secondary Alexa 488 goat anti-mouse antibody diluted 1:1,000 (Invitrogen), followed by addition of DAPI (4′,6-diamidino-2-phenylindole) to visualize cell nuclei. .. For some experiments, cells were visualized using an Axiovert 200 fluorescence microscope (Zeiss).

    Staining:

    Article Title: DDX56 Binds to Chikungunya Virus RNA To Control Infection
    Article Snippet: At the indicated time points, cells were washed once with phosphate-buffered saline (PBS) and collected in TRIzol (15596; Thermo) for RNA extraction according to the manufacturer’s instructions or fixed with 4% formaldehyde for 10 min for quantification by microscopy. .. Cells were stained for immunofluorescence imaging using anti-CHIKV ascites V-548-701-562 (ATCC VR-1241AF) at 1:300 or CHK-265 (gift from M. Diamond) at 1:500 and anti-mouse 488 and Hoechst 33342 (B2261; Sigma-Aldrich) at a final concentration of 5 μg/ml. ..

    Article Title: Chikungunya Virus Strains from Each Genetic Clade Bind Sulfated Glycosaminoglycans as Attachment Factors
    Article Snippet: .. Blocking buffer (PBS+/+ supplemented to contain 5% FBS and 0.1% TX-100) was added to the plate at room temperature and left for 1 h. Cells were stained with anti-CHIKV ascites fluid (1:1,500; ATCC VR-1241AF) at room temperature for 1 h and with goat anti-mouse Alexa Fluor 488 IgG (1:1,000; Invitrogen A11029 ) with 4′,6-diamidino-2-phenylindole (DAPI) (1:1,000; Thermo Fisher Scientific) at room temperature for 1 h. Cells were washed with PBS−/− three times at room temperature for 5 min per wash between each staining step. .. Infectivity was quantified by indirect immunofluorescence using the Lionheart FX automated microscope and Gen5 software (BioTek).

    Article Title: A Single-Amino-Acid Polymorphism in Chikungunya Virus E2 Glycoprotein Influences Glycosaminoglycan Utilization
    Article Snippet: Infected cells were visualized using indirect immunofluorescence. .. Cells were incubated in blocking buffer (phosphate-buffered saline [PBS; Gibco] containing 5% FBS and 0.1% Triton X-100) at room temperature for 1 h and stained with precleared anti-CHIKV immune ascetic fluid (ATCC VR-1241AF) diluted 1:1,500 and secondary Alexa 488 goat anti-mouse antibody diluted 1:1,000 (Invitrogen), followed by addition of DAPI (4′,6-diamidino-2-phenylindole) to visualize cell nuclei. .. For some experiments, cells were visualized using an Axiovert 200 fluorescence microscope (Zeiss).

    Immunofluorescence:

    Article Title: DDX56 Binds to Chikungunya Virus RNA To Control Infection
    Article Snippet: At the indicated time points, cells were washed once with phosphate-buffered saline (PBS) and collected in TRIzol (15596; Thermo) for RNA extraction according to the manufacturer’s instructions or fixed with 4% formaldehyde for 10 min for quantification by microscopy. .. Cells were stained for immunofluorescence imaging using anti-CHIKV ascites V-548-701-562 (ATCC VR-1241AF) at 1:300 or CHK-265 (gift from M. Diamond) at 1:500 and anti-mouse 488 and Hoechst 33342 (B2261; Sigma-Aldrich) at a final concentration of 5 μg/ml. ..

    Article Title: Next Generation Sequencing of DNA-Launched Chikungunya Vaccine Virus
    Article Snippet: To prepare virus for NGS, transfection supernatants were harvested at indicated times post-transfection and the virus titer was determined by standard plaque assay in Vero cells. .. Aliquots of transfected cell suspension were taken for CHIKV expression tests by immunofluorescence assay (IFA), infectious center assay (ICA) and western blot using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-64 (ATCC VR-1241AF) prepared against Chikungunya strain S-27. .. For ICA, electroporated cells were seeded into 6-well plates, allowed to adhere for 4 h, covered with 1% low-melting agarose in complete medium (aMEM containing 10% FBS and 10 μg/ml gentamicin), and incubated for 72 h. To visualize plaques, monolayers were stained using neutral red and counted the following day.

    Article Title: DNA Vaccine Initiates Replication of Live Attenuated Chikungunya Virus In Vitro and Elicits Protective Immune Response in Mice
    Article Snippet: As controls, cells were incubated with 102 –105 plaque-forming units (PFUs) of CHIKV 181/25 vaccine virus. .. Expression of CHIKV antigens in iDNA-transfected and virus-infected cells was detected by immunofluorescence assay (IFA) and Western blot, using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-1241AF (ATCC). .. CHIKV antigens were also confirmed by Western blot, using convalescent human antiserum (UTMB; courtesy of Dr Robert Tesh).

    Imaging:

    Article Title: DDX56 Binds to Chikungunya Virus RNA To Control Infection
    Article Snippet: At the indicated time points, cells were washed once with phosphate-buffered saline (PBS) and collected in TRIzol (15596; Thermo) for RNA extraction according to the manufacturer’s instructions or fixed with 4% formaldehyde for 10 min for quantification by microscopy. .. Cells were stained for immunofluorescence imaging using anti-CHIKV ascites V-548-701-562 (ATCC VR-1241AF) at 1:300 or CHK-265 (gift from M. Diamond) at 1:500 and anti-mouse 488 and Hoechst 33342 (B2261; Sigma-Aldrich) at a final concentration of 5 μg/ml. ..

    Concentration Assay:

    Article Title: DDX56 Binds to Chikungunya Virus RNA To Control Infection
    Article Snippet: At the indicated time points, cells were washed once with phosphate-buffered saline (PBS) and collected in TRIzol (15596; Thermo) for RNA extraction according to the manufacturer’s instructions or fixed with 4% formaldehyde for 10 min for quantification by microscopy. .. Cells were stained for immunofluorescence imaging using anti-CHIKV ascites V-548-701-562 (ATCC VR-1241AF) at 1:300 or CHK-265 (gift from M. Diamond) at 1:500 and anti-mouse 488 and Hoechst 33342 (B2261; Sigma-Aldrich) at a final concentration of 5 μg/ml. ..

    Blocking Assay:

    Article Title: Chikungunya Virus Strains from Each Genetic Clade Bind Sulfated Glycosaminoglycans as Attachment Factors
    Article Snippet: .. Blocking buffer (PBS+/+ supplemented to contain 5% FBS and 0.1% TX-100) was added to the plate at room temperature and left for 1 h. Cells were stained with anti-CHIKV ascites fluid (1:1,500; ATCC VR-1241AF) at room temperature for 1 h and with goat anti-mouse Alexa Fluor 488 IgG (1:1,000; Invitrogen A11029 ) with 4′,6-diamidino-2-phenylindole (DAPI) (1:1,000; Thermo Fisher Scientific) at room temperature for 1 h. Cells were washed with PBS−/− three times at room temperature for 5 min per wash between each staining step. .. Infectivity was quantified by indirect immunofluorescence using the Lionheart FX automated microscope and Gen5 software (BioTek).

    Article Title: A Single-Amino-Acid Polymorphism in Chikungunya Virus E2 Glycoprotein Influences Glycosaminoglycan Utilization
    Article Snippet: Infected cells were visualized using indirect immunofluorescence. .. Cells were incubated in blocking buffer (phosphate-buffered saline [PBS; Gibco] containing 5% FBS and 0.1% Triton X-100) at room temperature for 1 h and stained with precleared anti-CHIKV immune ascetic fluid (ATCC VR-1241AF) diluted 1:1,500 and secondary Alexa 488 goat anti-mouse antibody diluted 1:1,000 (Invitrogen), followed by addition of DAPI (4′,6-diamidino-2-phenylindole) to visualize cell nuclei. .. For some experiments, cells were visualized using an Axiovert 200 fluorescence microscope (Zeiss).

    Transfection:

    Article Title: Next Generation Sequencing of DNA-Launched Chikungunya Vaccine Virus
    Article Snippet: To prepare virus for NGS, transfection supernatants were harvested at indicated times post-transfection and the virus titer was determined by standard plaque assay in Vero cells. .. Aliquots of transfected cell suspension were taken for CHIKV expression tests by immunofluorescence assay (IFA), infectious center assay (ICA) and western blot using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-64 (ATCC VR-1241AF) prepared against Chikungunya strain S-27. .. For ICA, electroporated cells were seeded into 6-well plates, allowed to adhere for 4 h, covered with 1% low-melting agarose in complete medium (aMEM containing 10% FBS and 10 μg/ml gentamicin), and incubated for 72 h. To visualize plaques, monolayers were stained using neutral red and counted the following day.

    Expressing:

    Article Title: Next Generation Sequencing of DNA-Launched Chikungunya Vaccine Virus
    Article Snippet: To prepare virus for NGS, transfection supernatants were harvested at indicated times post-transfection and the virus titer was determined by standard plaque assay in Vero cells. .. Aliquots of transfected cell suspension were taken for CHIKV expression tests by immunofluorescence assay (IFA), infectious center assay (ICA) and western blot using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-64 (ATCC VR-1241AF) prepared against Chikungunya strain S-27. .. For ICA, electroporated cells were seeded into 6-well plates, allowed to adhere for 4 h, covered with 1% low-melting agarose in complete medium (aMEM containing 10% FBS and 10 μg/ml gentamicin), and incubated for 72 h. To visualize plaques, monolayers were stained using neutral red and counted the following day.

    Article Title: DNA Vaccine Initiates Replication of Live Attenuated Chikungunya Virus In Vitro and Elicits Protective Immune Response in Mice
    Article Snippet: As controls, cells were incubated with 102 –105 plaque-forming units (PFUs) of CHIKV 181/25 vaccine virus. .. Expression of CHIKV antigens in iDNA-transfected and virus-infected cells was detected by immunofluorescence assay (IFA) and Western blot, using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-1241AF (ATCC). .. CHIKV antigens were also confirmed by Western blot, using convalescent human antiserum (UTMB; courtesy of Dr Robert Tesh).

    Western Blot:

    Article Title: Next Generation Sequencing of DNA-Launched Chikungunya Vaccine Virus
    Article Snippet: To prepare virus for NGS, transfection supernatants were harvested at indicated times post-transfection and the virus titer was determined by standard plaque assay in Vero cells. .. Aliquots of transfected cell suspension were taken for CHIKV expression tests by immunofluorescence assay (IFA), infectious center assay (ICA) and western blot using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-64 (ATCC VR-1241AF) prepared against Chikungunya strain S-27. .. For ICA, electroporated cells were seeded into 6-well plates, allowed to adhere for 4 h, covered with 1% low-melting agarose in complete medium (aMEM containing 10% FBS and 10 μg/ml gentamicin), and incubated for 72 h. To visualize plaques, monolayers were stained using neutral red and counted the following day.

    Article Title: DNA Vaccine Initiates Replication of Live Attenuated Chikungunya Virus In Vitro and Elicits Protective Immune Response in Mice
    Article Snippet: As controls, cells were incubated with 102 –105 plaque-forming units (PFUs) of CHIKV 181/25 vaccine virus. .. Expression of CHIKV antigens in iDNA-transfected and virus-infected cells was detected by immunofluorescence assay (IFA) and Western blot, using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-1241AF (ATCC). .. CHIKV antigens were also confirmed by Western blot, using convalescent human antiserum (UTMB; courtesy of Dr Robert Tesh).

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  • 95
    ATCC chikv antigens
    Transfection of immunization DNA <t>(iDNA)</t> plasmid p181/25-7 into CHO cells. A , The immunization DNA (iDNA) approach for launching chikungunya virus <t>(CHIKV)</t> live attenuated virus in eukaryotic cells is shown on the left. Indicated are the cytomegalovirus (CMV) promoter (open arrow), cell nucleus, and progeny virus. Expression of CHIKV antigens after transfection of iDNA plasmid is shown on the right, detected by immunofluorescence assay (IFA) 48 and 96 hours after transfection. Aliquots of transfected cells were seeded in 8-well chamber slides, fixed at indicated times in cold acetone, and processed by IFA, using mouse CHIKV-specific antibody, followed by fluorescein isothiocyanate–conjugated secondary antibody. B , Detection of CHIKV antigens in transfected CHO cells by Western blot (left) and in the growth medium by plaque assay 48 hours after transfection (middle). For comparison, plaque assay for the 181/25 virus vaccine (passage 1 in CHO cells) is shown. Right panel shows growth curve of p181/25-7 iDNA-derived virus (average of 3 experiments). Western blot was done using human convalescent-phase CHIKV-specific serum (lane 1) and CHIKV HMAF (lane 2). The PE2, E2, E1, and C antigens are indicated.
    Chikv Antigens, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/chikv antigens/product/ATCC
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    chikv antigens - by Bioz Stars, 2021-07
    95/100 stars
      Buy from Supplier

    93
    ATCC chikv antigen
    Transfection of immunization DNA <t>(iDNA)</t> plasmid p181/25-7 into CHO cells. A , The immunization DNA (iDNA) approach for launching chikungunya virus <t>(CHIKV)</t> live attenuated virus in eukaryotic cells is shown on the left. Indicated are the cytomegalovirus (CMV) promoter (open arrow), cell nucleus, and progeny virus. Expression of CHIKV antigens after transfection of iDNA plasmid is shown on the right, detected by immunofluorescence assay (IFA) 48 and 96 hours after transfection. Aliquots of transfected cells were seeded in 8-well chamber slides, fixed at indicated times in cold acetone, and processed by IFA, using mouse CHIKV-specific antibody, followed by fluorescein isothiocyanate–conjugated secondary antibody. B , Detection of CHIKV antigens in transfected CHO cells by Western blot (left) and in the growth medium by plaque assay 48 hours after transfection (middle). For comparison, plaque assay for the 181/25 virus vaccine (passage 1 in CHO cells) is shown. Right panel shows growth curve of p181/25-7 iDNA-derived virus (average of 3 experiments). Western blot was done using human convalescent-phase CHIKV-specific serum (lane 1) and CHIKV HMAF (lane 2). The PE2, E2, E1, and C antigens are indicated.
    Chikv Antigen, supplied by ATCC, 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/chikv antigen/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    chikv antigen - by Bioz Stars, 2021-07
    93/100 stars
      Buy from Supplier

    Image Search Results


    Transfection of immunization DNA (iDNA) plasmid p181/25-7 into CHO cells. A , The immunization DNA (iDNA) approach for launching chikungunya virus (CHIKV) live attenuated virus in eukaryotic cells is shown on the left. Indicated are the cytomegalovirus (CMV) promoter (open arrow), cell nucleus, and progeny virus. Expression of CHIKV antigens after transfection of iDNA plasmid is shown on the right, detected by immunofluorescence assay (IFA) 48 and 96 hours after transfection. Aliquots of transfected cells were seeded in 8-well chamber slides, fixed at indicated times in cold acetone, and processed by IFA, using mouse CHIKV-specific antibody, followed by fluorescein isothiocyanate–conjugated secondary antibody. B , Detection of CHIKV antigens in transfected CHO cells by Western blot (left) and in the growth medium by plaque assay 48 hours after transfection (middle). For comparison, plaque assay for the 181/25 virus vaccine (passage 1 in CHO cells) is shown. Right panel shows growth curve of p181/25-7 iDNA-derived virus (average of 3 experiments). Western blot was done using human convalescent-phase CHIKV-specific serum (lane 1) and CHIKV HMAF (lane 2). The PE2, E2, E1, and C antigens are indicated.

    Journal: The Journal of Infectious Diseases

    Article Title: DNA Vaccine Initiates Replication of Live Attenuated Chikungunya Virus In Vitro and Elicits Protective Immune Response in Mice

    doi: 10.1093/infdis/jiu114

    Figure Lengend Snippet: Transfection of immunization DNA (iDNA) plasmid p181/25-7 into CHO cells. A , The immunization DNA (iDNA) approach for launching chikungunya virus (CHIKV) live attenuated virus in eukaryotic cells is shown on the left. Indicated are the cytomegalovirus (CMV) promoter (open arrow), cell nucleus, and progeny virus. Expression of CHIKV antigens after transfection of iDNA plasmid is shown on the right, detected by immunofluorescence assay (IFA) 48 and 96 hours after transfection. Aliquots of transfected cells were seeded in 8-well chamber slides, fixed at indicated times in cold acetone, and processed by IFA, using mouse CHIKV-specific antibody, followed by fluorescein isothiocyanate–conjugated secondary antibody. B , Detection of CHIKV antigens in transfected CHO cells by Western blot (left) and in the growth medium by plaque assay 48 hours after transfection (middle). For comparison, plaque assay for the 181/25 virus vaccine (passage 1 in CHO cells) is shown. Right panel shows growth curve of p181/25-7 iDNA-derived virus (average of 3 experiments). Western blot was done using human convalescent-phase CHIKV-specific serum (lane 1) and CHIKV HMAF (lane 2). The PE2, E2, E1, and C antigens are indicated.

    Article Snippet: Expression of CHIKV antigens in iDNA-transfected and virus-infected cells was detected by immunofluorescence assay (IFA) and Western blot, using CHIKV hyperimmune mouse ascitic fluid (HMAF) VR-1241AF (ATCC).

    Techniques: Transfection, Plasmid Preparation, Expressing, Immunofluorescence, Western Blot, Plaque Assay, Derivative Assay