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Mabtech Inc tmb elispot substrate solution
a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with <t>ELISpot.</t> Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.
Tmb Elispot Substrate Solution, supplied by Mabtech Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/elispot+substrate+solution/tmb+substrate+for+elispot/pmc10169129-346-4-8
Average 90 stars, based on 1 article reviews
tmb elispot substrate solution - by Bioz Stars, 2026-10
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1) Product Images from "Cationic crosslinked carbon dots-adjuvanted intranasal vaccine induces protective immunity against Omicron-included SARS-CoV-2 variants"

Article Title: Cationic crosslinked carbon dots-adjuvanted intranasal vaccine induces protective immunity against Omicron-included SARS-CoV-2 variants

Journal: Nature Communications

doi: 10.1038/s41467-023-38066-8

a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with ELISpot. Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.
Figure Legend Snippet: a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with ELISpot. Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.

Techniques Used: Variant Assay, Enzyme-linked Immunospot, Clinical Proteomics, Expressing, Staining

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Article Snippet: The next day, the plates were washed with PBS 4 times and incubated with 100 μl/well HRP-conjugated goat anti-mouse IgG antibody (1:10000, Cat: 0107-05, Southern Biotech) for 2 h at room temperature. .. After washing with PBS 4 times, TMB ELISpot substrate solution (100 μl/well, Cat:3651-10, Mabtech, Sweden) was added to the plates until a distinct spot emerged. ..

Article Title: Heterologous Prime-Boost with Immunologically Orthogonal Protein Nanoparticles for Peptide Immunofocusing
Article Snippet: The plates were then washed 5x times with PBST to remove cells and debris, then incubated at room temperature with biotinylated secondary goat antimouse IgG (catalog #3825-6-250, Mabtech) at a 2 μg/mL concentration in the dark for 2 h. Plates were washed 6× times with PBST and plated with streptavidin-HRP conjugate (1:1000 dilution; catalog #3310-9-1000, Mabtech) for 1 h at room temperature in the dark. .. The plates were then washed 3× with PBST, 3× with PBS, then followed by addition of 100 μL/well of ELISpot substrate (catalog #3651-10; Mabtech). ..

Article Title: The GP2a 91/97/98 amino acid substitutions play critical roles in determining PRRSV tropism and infectivity but do not affect immune responses.
Article Snippet: After 24 h incubation, the plates were washed and incubated with biotinylated anti-porcine IFN-γ mAb (0.5 μg/mL, P2C11, Mabtech, Sweden) for 2 h, followed by adding streptavidin-HRP (1:1,000, Mabtech, Sweden). .. Spots were developed with TMB ELISpot substrate (Mabtech, Sweden) and quantitated using an S6-Entry ImmunoSpot Analyzer (CTL, USA). ..

Article Title: Enhancing anti-viral neutralization response to immunization with HIV-1 envelope glycoprotein immunogens.
Article Snippet: .. ELISpot substrate (TMB; Mabtech, Cincinnati, OH) was added until distinct spots appeared. ..

Article Title: Heterologous Prime-Boost with Immunologically Orthogonal Protein Nanoparticles for Peptide Immunofocusing
Article Snippet: Plates were washed 6x times with PBST and plated with streptavidin-HRP conjugate (1:1000 dilution; catalogue #3310-9-1000, Mabtech) for 1 hour at room temperature in the dark. .. The plates were then washed 3x with PBST, 3x with PBS, then followed by addition of 100 μL/well of ELISpot substrate (catalogue #3651-10; Mabtech). ..

Article Title: Cellular immune signatures and differences of four porcine circovirus type 2 vaccines to heterologous PCV2d infection
Article Snippet: .. After 24-h incubation at 37 °C, the plates were washed and biotinylated anti‐porcine IFNγ mAb (0.5 μg/mL, P2C11, Mabtech, Sweden) was added, followed by streptavidin‐HRP (1:1000, Mabtech, Sweden), and spots were visualized using TMB ELISpot substrate (Mabtech, Sweden). .. Plates were dried and spots were counted on a S6-Entry ImmunoSpot Analyzer (CTL, USA).

Article Title: Comparison of a SARS-CoV-2 mRNA booster immunization containing additional antigens to a spike-based mRNA vaccine against Omicron BA.5 infection in hACE2 mice
Article Snippet: Following incubation, plates were washed with PBS and Streptavidin-horseradish peroxidase (HRP) enzyme conjugate (Mabtech) was added onto plates for 1 hour at room temperature. .. Plates were then washed again and room temperature TMB ELISpot substrate (Mabtech) was added onto plates and left until spots were visible. ..

Article Title: Heterologous Prime-Boost with Immunologically Orthogonal Protein Nanoparticles for Peptide Immunofocusing.
Article Snippet: The plates were then washed 5x times with PBST to remove cells and debris, then incubated at room temperature with biotinylated secondary goat antimouse IgG (catalog #3825-6-250, Mabtech) at a 2 μg/mL concentration in the dark for 2 h. Plates were washed 6× times with PBST and plated with streptavidin-HRP conjugate (1:1000 dilution; catalog #3310-9-1000, Mabtech) for 1 h at room temperature in the dark. .. The plates were then washed 3× with PBST, 3×with PBS, then followed by addition of 100 μL/well of ELISpot substrate (catalog #3651-10; Mabtech). ..

Incubation:

Article Title: Cellular immune signatures and differences of four porcine circovirus type 2 vaccines to heterologous PCV2d infection
Article Snippet: .. After 24-h incubation at 37 °C, the plates were washed and biotinylated anti‐porcine IFNγ mAb (0.5 μg/mL, P2C11, Mabtech, Sweden) was added, followed by streptavidin‐HRP (1:1000, Mabtech, Sweden), and spots were visualized using TMB ELISpot substrate (Mabtech, Sweden). .. Plates were dried and spots were counted on a S6-Entry ImmunoSpot Analyzer (CTL, USA).



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a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with <t>ELISpot.</t> Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.
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Image Search Results


a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with ELISpot. Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Cationic crosslinked carbon dots-adjuvanted intranasal vaccine induces protective immunity against Omicron-included SARS-CoV-2 variants

doi: 10.1038/s41467-023-38066-8

Figure Lengend Snippet: a Immunization scheme. Mice/rabbits were immunized intranasally with PBS, RBD-HR, or CCD/RBD-HR on days 0, 14, and 28. b , c Serum anti-RBD-HR IgG titers in immunized mice ( b ) and rabbits ( c ) on day 35. d , e Neutralizing of authentic viruses ( d ) and pseudoviruses ( e ) of SARS-CoV-2 by the immune sera on day 35. f RBD-HR-specific IgA levels in BAL fluid on day 35. Immunized mice were challenged with 5 × 10 5 TCID 50 of Omicron viruses and sacrificed at 4 dpi. g Viral RNA levels in the nasal turbinates, trachea, and lungs at 4 dpi. h Histopathological changes (left) and pathological scores (right) of the lung tissues at 4 dpi. Scale bar: 100 μm. Similar results were obtained in five independent experiments. i, j Serum RBD-HR-specific IgG levels ( i ) and their function in neutralizing SARS-CoV-2 variant pseudoviruses ( j ) on day 260. On day 56, blood ( k ), bone marrow ( l ), and spleen ( m ) were harvested to assess SARS-CoV-2 RBD-specific IgG ASCs with ELISpot. Images (left) and quantification (right) of IgG spot-forming cells among lymphocytes were displayed. RBD-specific IgG-producing CD138 + plasma cells in the blood and bone marrow ( n ), memory B cells (RBD + CD38 + CD80 + ) in the blood, bone marrow, and spleen ( o ) were assayed with FCM. p , q Antigen-experienced (CD44 + ) and activated (CD69 + ) CD4 + /CD8 + T cells in the spleen were determined with FCM after re-stimulation with RBD protein peptide pools. r Splenic CD4 + and CD8 + T cells were assessed for IL-4 and IFN-γ expression with intracellular cytokine staining after restimulation with RBD protein peptide pools or irrelevant antigens. Data were displayed with floating bars in ( k – r ). The middle line indicates the median and the box shows the data range. Data were presented as mean values ± SEM. n = 4–5 mice in ( b , d – r ). n = 3 rabbits in ( c ). P values were calculated with one-way ANOVA followed by Dunnett’s multiple comparisons test in ( b , c , f – i , k – q) . Two-way ANOVA followed by Sidak’s multiple comparisons test in ( d , e , and j ). Two-way ANOVA followed by Tukey’s multiple comparisons test in ( r ). Source data are provided as a Source Data file.

Article Snippet: Following additional five washes, TMB ELISpot substrate solution (Mabtech, Sweden) was added for spot formation.

Techniques: Variant Assay, Enzyme-linked Immunospot, Clinical Proteomics, Expressing, Staining

IFN-γ ELISpot assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), or PBS (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.

Journal: Heliyon

Article Title: Solar inactivated Salmonella Typhimurium induces an immune response in BALB/c mice

doi: 10.1016/j.heliyon.2021.e05903

Figure Lengend Snippet: IFN-γ ELISpot assay showing the number of spot forming splenocytes in mice administered with either solar inactivated S . Typhimurium (green), or PBS (blue). LPS, FliC and BACT, were used as stimulants. Each data point (circle or square) represents an individual mouse, (from 1 experiment (Day 42)) and the error bars denote the interquartile range of the median. Data was analysed with a 2-way analysis of variance and post-hoc positive test, p values indicate significant differences between treatments groups for individual antigens.

Article Snippet: Following incubation for at least 1 h at room temperature and washing six times with PBS, ELISpot colour development solution (170-6432, Bio-Rad, CA) was added.

Techniques: Enzyme-linked Immunospot