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elispot flex mouse ifn γ alp kit  (Mabtech Inc)

 
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    Mabtech Inc elispot flex mouse ifn γ alp kit
    Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ <t>ELISpot</t> analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.
    Elispot Flex Mouse Ifn γ Alp Kit, supplied by Mabtech Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/elispot+flex+mouse+ifn+%CE%B3+alp+kit/antibody+biotinylated+ifn%CE%B3+specific/pmc13020013-181-13-19
    Average 86 stars, based on 1 article reviews
    elispot flex mouse ifn γ alp kit - by Bioz Stars, 2026-11
    86/100 stars

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    1) Product Images from "Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses"

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses

    Journal: eBioMedicine

    doi: 10.1016/j.ebiom.2026.106228

    Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.
    Figure Legend Snippet: Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.

    Techniques Used: Mutagenesis, RNA Binding Assay, Produced, Fluorescence, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Generated, MANN-WHITNEY

    Mouse antibody and T cell responses following RSPVac nasal vaccination. (A) Vaccination regimen. Female BALB/c mice (n = 6–9) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Sera, BALF, and BALF cells were harvested at day 10 post-2nd dose for analysis. Control mice received the protein component without RNA (protein-only). Data was shown as box plots with respective colours indicating vaccine given, showing all values. Error bars represented the highest and lowest values of each box. ELISA was used to measure antibody responses for (B) anti-HA serum IgG, (C) anti-HA BALF IgG, (D) anti-HA BALF IgA, (E) anti-NP, and (F) serum anti-HA IgG subtypes for each of the RSPVac, respectively. (G) Mucosal T cell responses measured by IFNγ+ ELISpot using the immunogen (the protein used to generate RSPVac) stimulation of BALF cells. (H) To analyse lung T cells, female BALB/c mice (n = 6) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Lungs were harvested at day 7 post-2nd dose. Single cell suspensions were prepared, and the NP peptide pool of H5N1, H1N1, and H7N9 was used to stimulate lung cells. Resident T cell responses were analysed by intracellular staining and FACS analysis. NP-reactive (I) CD4+ and (J) CD8+ T cells were shown, comparing protein-only and RSPVac-vaccinated. Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered insignificant were labelled not significant (ns). Figures A and H were created in BioRender.
    Figure Legend Snippet: Mouse antibody and T cell responses following RSPVac nasal vaccination. (A) Vaccination regimen. Female BALB/c mice (n = 6–9) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Sera, BALF, and BALF cells were harvested at day 10 post-2nd dose for analysis. Control mice received the protein component without RNA (protein-only). Data was shown as box plots with respective colours indicating vaccine given, showing all values. Error bars represented the highest and lowest values of each box. ELISA was used to measure antibody responses for (B) anti-HA serum IgG, (C) anti-HA BALF IgG, (D) anti-HA BALF IgA, (E) anti-NP, and (F) serum anti-HA IgG subtypes for each of the RSPVac, respectively. (G) Mucosal T cell responses measured by IFNγ+ ELISpot using the immunogen (the protein used to generate RSPVac) stimulation of BALF cells. (H) To analyse lung T cells, female BALB/c mice (n = 6) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Lungs were harvested at day 7 post-2nd dose. Single cell suspensions were prepared, and the NP peptide pool of H5N1, H1N1, and H7N9 was used to stimulate lung cells. Resident T cell responses were analysed by intracellular staining and FACS analysis. NP-reactive (I) CD4+ and (J) CD8+ T cells were shown, comparing protein-only and RSPVac-vaccinated. Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered insignificant were labelled not significant (ns). Figures A and H were created in BioRender.

    Techniques Used: Control, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Single Cell, Staining, MANN-WHITNEY

    Related Articles

    Recombinant:

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses
    Article Snippet: After staining, cells were washed, passed through a 70 μm cell strainer, and analysed by BD Fortessa X-20 SORP cell analyser. .. BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A). .. BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A).

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses.
    Article Snippet: .. BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A). .. 1 × 10 4 BALF cells were cultured in activation medium (described above) on Multiscreen® 96 well mixed cellulose esters filter plate (Millipore, MAHAS4510) and stimulated with 10 μg of corresponding immunogen (the protein used to generate RSPVac) for 48 h. Plates were then washed, stained with kit antibodies, and spots were developed by Sigmafast BCIP/NBT (Sigma, B5655-25TAB) and counted by Cytation 7 (Biotek) or S6 Universal M2 (ImmunoSpot).

    Enzyme-linked Immunospot:

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses
    Article Snippet: After staining, cells were washed, passed through a 70 μm cell strainer, and analysed by BD Fortessa X-20 SORP cell analyser. .. BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A). .. BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A).

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses.
    Article Snippet: .. BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A). .. 1 × 10 4 BALF cells were cultured in activation medium (described above) on Multiscreen® 96 well mixed cellulose esters filter plate (Millipore, MAHAS4510) and stimulated with 10 μg of corresponding immunogen (the protein used to generate RSPVac) for 48 h. Plates were then washed, stained with kit antibodies, and spots were developed by Sigmafast BCIP/NBT (Sigma, B5655-25TAB) and counted by Cytation 7 (Biotek) or S6 Universal M2 (ImmunoSpot).



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    Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ <t>ELISpot</t> analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.
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    Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ <t>ELISpot</t> analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.
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    Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.

    Journal: eBioMedicine

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses

    doi: 10.1016/j.ebiom.2026.106228

    Figure Lengend Snippet: Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.

    Article Snippet: BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A).

    Techniques: Mutagenesis, RNA Binding Assay, Produced, Fluorescence, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Generated, MANN-WHITNEY

    Mouse antibody and T cell responses following RSPVac nasal vaccination. (A) Vaccination regimen. Female BALB/c mice (n = 6–9) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Sera, BALF, and BALF cells were harvested at day 10 post-2nd dose for analysis. Control mice received the protein component without RNA (protein-only). Data was shown as box plots with respective colours indicating vaccine given, showing all values. Error bars represented the highest and lowest values of each box. ELISA was used to measure antibody responses for (B) anti-HA serum IgG, (C) anti-HA BALF IgG, (D) anti-HA BALF IgA, (E) anti-NP, and (F) serum anti-HA IgG subtypes for each of the RSPVac, respectively. (G) Mucosal T cell responses measured by IFNγ+ ELISpot using the immunogen (the protein used to generate RSPVac) stimulation of BALF cells. (H) To analyse lung T cells, female BALB/c mice (n = 6) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Lungs were harvested at day 7 post-2nd dose. Single cell suspensions were prepared, and the NP peptide pool of H5N1, H1N1, and H7N9 was used to stimulate lung cells. Resident T cell responses were analysed by intracellular staining and FACS analysis. NP-reactive (I) CD4+ and (J) CD8+ T cells were shown, comparing protein-only and RSPVac-vaccinated. Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered insignificant were labelled not significant (ns). Figures A and H were created in BioRender.

    Journal: eBioMedicine

    Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses

    doi: 10.1016/j.ebiom.2026.106228

    Figure Lengend Snippet: Mouse antibody and T cell responses following RSPVac nasal vaccination. (A) Vaccination regimen. Female BALB/c mice (n = 6–9) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Sera, BALF, and BALF cells were harvested at day 10 post-2nd dose for analysis. Control mice received the protein component without RNA (protein-only). Data was shown as box plots with respective colours indicating vaccine given, showing all values. Error bars represented the highest and lowest values of each box. ELISA was used to measure antibody responses for (B) anti-HA serum IgG, (C) anti-HA BALF IgG, (D) anti-HA BALF IgA, (E) anti-NP, and (F) serum anti-HA IgG subtypes for each of the RSPVac, respectively. (G) Mucosal T cell responses measured by IFNγ+ ELISpot using the immunogen (the protein used to generate RSPVac) stimulation of BALF cells. (H) To analyse lung T cells, female BALB/c mice (n = 6) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Lungs were harvested at day 7 post-2nd dose. Single cell suspensions were prepared, and the NP peptide pool of H5N1, H1N1, and H7N9 was used to stimulate lung cells. Resident T cell responses were analysed by intracellular staining and FACS analysis. NP-reactive (I) CD4+ and (J) CD8+ T cells were shown, comparing protein-only and RSPVac-vaccinated. Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered insignificant were labelled not significant (ns). Figures A and H were created in BioRender.

    Article Snippet: BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A).

    Techniques: Control, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Single Cell, Staining, MANN-WHITNEY