pierce immunoprecipitation lysis buffer  (Thermo Fisher)


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    Structured Review

    Thermo Fisher pierce immunoprecipitation lysis buffer
    Effects of the amino acids at positions 286 and 437 of NP on NP-NP homo-oligomerization. (A) Schematic representation of influenza virus NP. (B and C) NP-NP homo-oligomerization assay results. NP-NP homo-oligomerization analysis was performed by transfecting 293T cells with Myc-tagged NP and Flag-tagged NP; at 36 h posttransfection, cell lysates were immunoprecipitated with an anti-Myc MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (B), or the cell lysates were immunoprecipitated with an anti-Flag MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (C). IP, <t>immunoprecipitation.</t> The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in panels B and C.
    Pierce Immunoprecipitation Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Amino Acid Mutations A286V and T437M in the Nucleoprotein Attenuate H7N9 Viruses in Mice"

    Article Title: Amino Acid Mutations A286V and T437M in the Nucleoprotein Attenuate H7N9 Viruses in Mice

    Journal: Journal of Virology

    doi: 10.1128/JVI.01530-19

    Effects of the amino acids at positions 286 and 437 of NP on NP-NP homo-oligomerization. (A) Schematic representation of influenza virus NP. (B and C) NP-NP homo-oligomerization assay results. NP-NP homo-oligomerization analysis was performed by transfecting 293T cells with Myc-tagged NP and Flag-tagged NP; at 36 h posttransfection, cell lysates were immunoprecipitated with an anti-Myc MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (B), or the cell lysates were immunoprecipitated with an anti-Flag MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (C). IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in panels B and C.
    Figure Legend Snippet: Effects of the amino acids at positions 286 and 437 of NP on NP-NP homo-oligomerization. (A) Schematic representation of influenza virus NP. (B and C) NP-NP homo-oligomerization assay results. NP-NP homo-oligomerization analysis was performed by transfecting 293T cells with Myc-tagged NP and Flag-tagged NP; at 36 h posttransfection, cell lysates were immunoprecipitated with an anti-Myc MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (B), or the cell lysates were immunoprecipitated with an anti-Flag MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (C). IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in panels B and C.

    Techniques Used: Immunoprecipitation, Western Blot, FLAG-tag, Software, Transfection

    Effects of the amino acid mutations A286V and T437M in NP on the interactions between NP and members of the importin α family. 293T cells were transfected with plasmids expressing Flag-tagged importin α1 (A), importin α3 (B), importin α5 (C), or importin α7 (D) and the indicated Myc-tagged wild-type NP or NP mutants. IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in the panels.
    Figure Legend Snippet: Effects of the amino acid mutations A286V and T437M in NP on the interactions between NP and members of the importin α family. 293T cells were transfected with plasmids expressing Flag-tagged importin α1 (A), importin α3 (B), importin α5 (C), or importin α7 (D) and the indicated Myc-tagged wild-type NP or NP mutants. IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in the panels.

    Techniques Used: Transfection, Expressing, Immunoprecipitation, Western Blot, Software

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    Thermo Fisher pierce immunoprecipitation lysis buffer
    Effects of the amino acids at positions 286 and 437 of NP on NP-NP homo-oligomerization. (A) Schematic representation of influenza virus NP. (B and C) NP-NP homo-oligomerization assay results. NP-NP homo-oligomerization analysis was performed by transfecting 293T cells with Myc-tagged NP and Flag-tagged NP; at 36 h posttransfection, cell lysates were immunoprecipitated with an anti-Myc MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (B), or the cell lysates were immunoprecipitated with an anti-Flag MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (C). IP, <t>immunoprecipitation.</t> The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in panels B and C.
    Pierce Immunoprecipitation Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher b 27
    DEX reduced microglia activation and attenuated neuronal cell apoptosis in vitro . DEX reduces microglia activation and attenuates neuronal cell apoptosis in vitro . The primary microglia viability and toxicity test were examined by the CCK-8 kit. Each group had three replicates, and the test was repeated 4 times independently (A) . LPS-primed (100 ng/ml) primary microglia were treated with different doses of DEX for 5 h and then stimulated with ATP (5 mM) for 30 min. The expression of IBA-1 detected by Immunoblotting (B) and the statistical data shown in (C) (means ± SEM, n = 4). LPS-primed primary microglial cells were incubated with DEX for 5 h, then stimulated with ATP for 30 min.Microglia conditioned medium including without any treatment, treated with LPS (100 ng/ml, 5 h)plus ATP (5 mM, 30 min), and DEX (10 μM, 5 h) protected, MCM mixed with 2% <t>B-27</t> Neurobasal medium (1:1) and incubated the cells for 24 h, primary neuronal cells were incubated with mix MCM separately, collected for Immunoblotting, immunofluorescence staining and cell flow analysis. Expression of apoptosis-related proteins BAX and Bcl-2 detected by Immunoblotting (D) , the statistical data are shown in (E,F) (means ± SEM, n = 4). Apoptosis indicators were detected using a cell flow cytometer (G) . Data analysis is shown in (H) (means ± SEM,n = 4). MAP-2 expression was stained by immunofluorescence and axon length was calculated by IMageJ software (I) , data analysis in (J) (means ± SEM, n = 4). All the above data analysis were presented as the means ± SEM,* p
    B 27, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher b per reagent
    E. piscicida 1906I induced robust membrane-ruptured cell death and showed attenuated bacterial colonization and virulence in zebrafish. (A) Dynamic changes of HeLa cell morphology after infection of wild-type EIB202 or mutant 1906I at an MOI of 25 for the indicated time. White arrows denote the membrane bebbling and black arrows denote membrane swelling and rounding. Propidium iodide (PI) was added to detect the loss of plasma membrane integrity. Scale bar, 20 μm. <t>(B)</t> Lactate dehydrogenase (LDH) release of HeLa cells infected with EIB202 or 1906I for the indicated time. (C–G) Adult zebrafish were intramuscular injected with 50 CFU indicated E. piscicida strains <t>per</t> fish. (C) Survival of zebrafish was monitored for 6 days. PBS as control. ( n =35). (D–G) Bacteria colonization in liver (D) , spleen (E) , kidney (F) and intestine (G) of the indicated time points post infection. Five surviving adult zebrafish were pooled as a sample and dissected at the indicated time point post infection, and livers, spleens, kidneys, and intestines were collected and grinded to homogenates for CFU counts, respectively. Results are representative of at least three independent experiments. * P
    B Per Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher buffer b
    Assessment of coupling efficiency using a dilution series of goat anti-GST IgG antibody. Goat anti-GST IgG antibody was diluted 1:1 × 10 3 , 1:1 × 10 4 , 1:5 × 10 4 , 1:1 × 10 5 , 1:5 × 10 5 , 1:1 × 10 6 , 1:5 × 10 6 , and 1:1 × 10 7  in modified Buffer A lacking the  E. coli  extract. Following a 1-h incubation (50 μl/well) at room temperature, bound anti-GST antibody was detected with a biotinylated rabbit anti-goat IgG secondary antibody (1:500 dilution in Buffer B, 50 μl/well, 1 h at room temperature). Wells were developed with  R -phycoerythrin-labelled streptavidin and read on a BioPlex 200 instrument (BioRad) as described in “  Methods ”
    Buffer B, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of the amino acids at positions 286 and 437 of NP on NP-NP homo-oligomerization. (A) Schematic representation of influenza virus NP. (B and C) NP-NP homo-oligomerization assay results. NP-NP homo-oligomerization analysis was performed by transfecting 293T cells with Myc-tagged NP and Flag-tagged NP; at 36 h posttransfection, cell lysates were immunoprecipitated with an anti-Myc MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (B), or the cell lysates were immunoprecipitated with an anti-Flag MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (C). IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in panels B and C.

    Journal: Journal of Virology

    Article Title: Amino Acid Mutations A286V and T437M in the Nucleoprotein Attenuate H7N9 Viruses in Mice

    doi: 10.1128/JVI.01530-19

    Figure Lengend Snippet: Effects of the amino acids at positions 286 and 437 of NP on NP-NP homo-oligomerization. (A) Schematic representation of influenza virus NP. (B and C) NP-NP homo-oligomerization assay results. NP-NP homo-oligomerization analysis was performed by transfecting 293T cells with Myc-tagged NP and Flag-tagged NP; at 36 h posttransfection, cell lysates were immunoprecipitated with an anti-Myc MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (B), or the cell lysates were immunoprecipitated with an anti-Flag MAb, followed by Western blotting with MAbs against the Myc tag and the Flag tag (C). IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in panels B and C.

    Article Snippet: Briefly, 293T cells were washed twice with cold PBS, lysed with Pierce immunoprecipitation lysis buffer (Invitrogen) containing complete protease inhibitor cocktail (Roche Diagnostics GmbH, Mannheim, Germany) for 30 min on ice, and then centrifuged at 12,000 rpm at 4°C for 10 min.

    Techniques: Immunoprecipitation, Western Blot, FLAG-tag, Software, Transfection

    Effects of the amino acid mutations A286V and T437M in NP on the interactions between NP and members of the importin α family. 293T cells were transfected with plasmids expressing Flag-tagged importin α1 (A), importin α3 (B), importin α5 (C), or importin α7 (D) and the indicated Myc-tagged wild-type NP or NP mutants. IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in the panels.

    Journal: Journal of Virology

    Article Title: Amino Acid Mutations A286V and T437M in the Nucleoprotein Attenuate H7N9 Viruses in Mice

    doi: 10.1128/JVI.01530-19

    Figure Lengend Snippet: Effects of the amino acid mutations A286V and T437M in NP on the interactions between NP and members of the importin α family. 293T cells were transfected with plasmids expressing Flag-tagged importin α1 (A), importin α3 (B), importin α5 (C), or importin α7 (D) and the indicated Myc-tagged wild-type NP or NP mutants. IP, immunoprecipitation. The band intensities of the Western blots from three assays were quantified by using ImageJ software and compared with the value of the CK/S1220-NP-transfected sample. Statistical analysis was performed by using one-way ANOVA with GraphPad Prism 6 software. When one-way ANOVA was warranted, the post hoc analysis was performed using Dunnett’s test for multiple comparisons; no statistically significant differences were detected. Only the band intensity values of the coimmunoprecipitated proteins are shown in the panels.

    Article Snippet: Briefly, 293T cells were washed twice with cold PBS, lysed with Pierce immunoprecipitation lysis buffer (Invitrogen) containing complete protease inhibitor cocktail (Roche Diagnostics GmbH, Mannheim, Germany) for 30 min on ice, and then centrifuged at 12,000 rpm at 4°C for 10 min.

    Techniques: Transfection, Expressing, Immunoprecipitation, Western Blot, Software

    DEX reduced microglia activation and attenuated neuronal cell apoptosis in vitro . DEX reduces microglia activation and attenuates neuronal cell apoptosis in vitro . The primary microglia viability and toxicity test were examined by the CCK-8 kit. Each group had three replicates, and the test was repeated 4 times independently (A) . LPS-primed (100 ng/ml) primary microglia were treated with different doses of DEX for 5 h and then stimulated with ATP (5 mM) for 30 min. The expression of IBA-1 detected by Immunoblotting (B) and the statistical data shown in (C) (means ± SEM, n = 4). LPS-primed primary microglial cells were incubated with DEX for 5 h, then stimulated with ATP for 30 min.Microglia conditioned medium including without any treatment, treated with LPS (100 ng/ml, 5 h)plus ATP (5 mM, 30 min), and DEX (10 μM, 5 h) protected, MCM mixed with 2% B-27 Neurobasal medium (1:1) and incubated the cells for 24 h, primary neuronal cells were incubated with mix MCM separately, collected for Immunoblotting, immunofluorescence staining and cell flow analysis. Expression of apoptosis-related proteins BAX and Bcl-2 detected by Immunoblotting (D) , the statistical data are shown in (E,F) (means ± SEM, n = 4). Apoptosis indicators were detected using a cell flow cytometer (G) . Data analysis is shown in (H) (means ± SEM,n = 4). MAP-2 expression was stained by immunofluorescence and axon length was calculated by IMageJ software (I) , data analysis in (J) (means ± SEM, n = 4). All the above data analysis were presented as the means ± SEM,* p

    Journal: Frontiers in Pharmacology

    Article Title: Dexmedetomidine Mitigated NLRP3-Mediated Neuroinflammation via the Ubiquitin-Autophagy Pathway to Improve Perioperative Neurocognitive Disorder in Mice

    doi: 10.3389/fphar.2021.646265

    Figure Lengend Snippet: DEX reduced microglia activation and attenuated neuronal cell apoptosis in vitro . DEX reduces microglia activation and attenuates neuronal cell apoptosis in vitro . The primary microglia viability and toxicity test were examined by the CCK-8 kit. Each group had three replicates, and the test was repeated 4 times independently (A) . LPS-primed (100 ng/ml) primary microglia were treated with different doses of DEX for 5 h and then stimulated with ATP (5 mM) for 30 min. The expression of IBA-1 detected by Immunoblotting (B) and the statistical data shown in (C) (means ± SEM, n = 4). LPS-primed primary microglial cells were incubated with DEX for 5 h, then stimulated with ATP for 30 min.Microglia conditioned medium including without any treatment, treated with LPS (100 ng/ml, 5 h)plus ATP (5 mM, 30 min), and DEX (10 μM, 5 h) protected, MCM mixed with 2% B-27 Neurobasal medium (1:1) and incubated the cells for 24 h, primary neuronal cells were incubated with mix MCM separately, collected for Immunoblotting, immunofluorescence staining and cell flow analysis. Expression of apoptosis-related proteins BAX and Bcl-2 detected by Immunoblotting (D) , the statistical data are shown in (E,F) (means ± SEM, n = 4). Apoptosis indicators were detected using a cell flow cytometer (G) . Data analysis is shown in (H) (means ± SEM,n = 4). MAP-2 expression was stained by immunofluorescence and axon length was calculated by IMageJ software (I) , data analysis in (J) (means ± SEM, n = 4). All the above data analysis were presented as the means ± SEM,* p

    Article Snippet: The cell pellet was resuspended in neurobasal medium (Gibco, Thermo Fisher Scientific) supplemented with 2% B-27 (Gibco, Thermo Fisher Scientific) and 0.5 mM L -glutamic acid.

    Techniques: Activation Assay, In Vitro, CCK-8 Assay, Expressing, Incubation, Immunofluorescence, Staining, Flow Cytometry, Software

    E. piscicida 1906I induced robust membrane-ruptured cell death and showed attenuated bacterial colonization and virulence in zebrafish. (A) Dynamic changes of HeLa cell morphology after infection of wild-type EIB202 or mutant 1906I at an MOI of 25 for the indicated time. White arrows denote the membrane bebbling and black arrows denote membrane swelling and rounding. Propidium iodide (PI) was added to detect the loss of plasma membrane integrity. Scale bar, 20 μm. (B) Lactate dehydrogenase (LDH) release of HeLa cells infected with EIB202 or 1906I for the indicated time. (C–G) Adult zebrafish were intramuscular injected with 50 CFU indicated E. piscicida strains per fish. (C) Survival of zebrafish was monitored for 6 days. PBS as control. ( n =35). (D–G) Bacteria colonization in liver (D) , spleen (E) , kidney (F) and intestine (G) of the indicated time points post infection. Five surviving adult zebrafish were pooled as a sample and dissected at the indicated time point post infection, and livers, spleens, kidneys, and intestines were collected and grinded to homogenates for CFU counts, respectively. Results are representative of at least three independent experiments. * P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Dysregulation of Cytosolic c-di-GMP in Edwardsiella piscicida Promotes Cellular Non-Canonical Ferroptosis

    doi: 10.3389/fcimb.2022.825824

    Figure Lengend Snippet: E. piscicida 1906I induced robust membrane-ruptured cell death and showed attenuated bacterial colonization and virulence in zebrafish. (A) Dynamic changes of HeLa cell morphology after infection of wild-type EIB202 or mutant 1906I at an MOI of 25 for the indicated time. White arrows denote the membrane bebbling and black arrows denote membrane swelling and rounding. Propidium iodide (PI) was added to detect the loss of plasma membrane integrity. Scale bar, 20 μm. (B) Lactate dehydrogenase (LDH) release of HeLa cells infected with EIB202 or 1906I for the indicated time. (C–G) Adult zebrafish were intramuscular injected with 50 CFU indicated E. piscicida strains per fish. (C) Survival of zebrafish was monitored for 6 days. PBS as control. ( n =35). (D–G) Bacteria colonization in liver (D) , spleen (E) , kidney (F) and intestine (G) of the indicated time points post infection. Five surviving adult zebrafish were pooled as a sample and dissected at the indicated time point post infection, and livers, spleens, kidneys, and intestines were collected and grinded to homogenates for CFU counts, respectively. Results are representative of at least three independent experiments. * P

    Article Snippet: 8×109 bacterium were collected at final as for each strain and 500 μl B-PER reagent (Thermo Fisher) was added to and mixed sufficiently with bacteria by pipetting.

    Techniques: Infection, Mutagenesis, Injection, Fluorescence In Situ Hybridization

    Assessment of coupling efficiency using a dilution series of goat anti-GST IgG antibody. Goat anti-GST IgG antibody was diluted 1:1 × 10 3 , 1:1 × 10 4 , 1:5 × 10 4 , 1:1 × 10 5 , 1:5 × 10 5 , 1:1 × 10 6 , 1:5 × 10 6 , and 1:1 × 10 7  in modified Buffer A lacking the  E. coli  extract. Following a 1-h incubation (50 μl/well) at room temperature, bound anti-GST antibody was detected with a biotinylated rabbit anti-goat IgG secondary antibody (1:500 dilution in Buffer B, 50 μl/well, 1 h at room temperature). Wells were developed with  R -phycoerythrin-labelled streptavidin and read on a BioPlex 200 instrument (BioRad) as described in “  Methods ”

    Journal: Malaria Journal

    Article Title: Specificity of the IgG antibody response to Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale MSP119 subunit proteins in multiplexed serologic assays

    doi: 10.1186/s12936-018-2566-0

    Figure Lengend Snippet: Assessment of coupling efficiency using a dilution series of goat anti-GST IgG antibody. Goat anti-GST IgG antibody was diluted 1:1 × 10 3 , 1:1 × 10 4 , 1:5 × 10 4 , 1:1 × 10 5 , 1:5 × 10 5 , 1:1 × 10 6 , 1:5 × 10 6 , and 1:1 × 10 7 in modified Buffer A lacking the E. coli extract. Following a 1-h incubation (50 μl/well) at room temperature, bound anti-GST antibody was detected with a biotinylated rabbit anti-goat IgG secondary antibody (1:500 dilution in Buffer B, 50 μl/well, 1 h at room temperature). Wells were developed with R -phycoerythrin-labelled streptavidin and read on a BioPlex 200 instrument (BioRad) as described in “ Methods ”

    Article Snippet: Washed beads were then incubated for 45 min at room temperature with 50 μl/well of a 1:500 dilution in Buffer B (0.5% BSA, 0.05% Tween-20, and 0.02% NaN3 in PBS at pH 7.2) of biotinylated monoclonal mouse anti-human IgG subclass secondary antibody to IgG1 (clone HP6025), IgG2 (clone HP6002), IgG3 (clone HP6047), or IgG4 (clone HP6025) (all from Zymed/Invitrogen, South San Francisco, CA, USA).

    Techniques: Modification, Incubation