mtt assay Search Results


rna extraction  (ATCC)
96
ATCC rna extraction
Rna Extraction, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mtt assay 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide  (Gold Biotechnology Inc)
95
Gold Biotechnology Inc mtt assay 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Mtt Assay 3 4 5 Dimethylthiazol 2 Yl 2 5 Diphenyltetrazolium Bromide, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mtt solution 3  (Merck & Co)
86
Merck & Co mtt solution 3
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Mtt Solution 3, supplied by Merck & Co, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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methylthiazolyldiphenyl tetrazolium  (MedChemExpress)
95
MedChemExpress methylthiazolyldiphenyl tetrazolium
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Methylthiazolyldiphenyl Tetrazolium, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cytotoxicity assay kit  (Beyotime)
96
Beyotime cytotoxicity assay kit
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Cytotoxicity Assay Kit, supplied by Beyotime, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mtt cell proliferation viability assay kit  (R&D Systems)
94
R&D Systems mtt cell proliferation viability assay kit
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Mtt Cell Proliferation Viability Assay Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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tacs mtt cell proliferation assay  (R&D Systems)
94
R&D Systems tacs mtt cell proliferation assay
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Tacs Mtt Cell Proliferation Assay, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mtt solution  (Beijing Solarbio Science)
96
Beijing Solarbio Science mtt solution
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Mtt Solution, supplied by Beijing Solarbio Science, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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5224 tocris brand  (Tocris)
93
Tocris 5224 tocris brand
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
5224 Tocris Brand, supplied by Tocris, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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4890 050 k  (R&D Systems)
94
R&D Systems 4890 050 k
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
4890 050 K, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mtt reagent  (R&D Systems)
93
R&D Systems mtt reagent
( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by <t>MTT</t> <t>assay</t> at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Mtt Reagent, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by MTT assay at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Journal: bioRxiv

Article Title: Integrating Fas-mediated apoptosis with IFNγ signaling to drive tumor regression in mRNA cancer therapeutics

doi: 10.64898/2026.04.06.716844

Figure Lengend Snippet: ( A ) Schematic diagram of IFNγ-FasICD and IFNγ-FasΔ proteins. SP; The IFNγ signal sequence peptide, FasTM; Fas transmembrane domain, FasICD; Fas intracellular domain. A linker (GGGSGGGSGGGS) was added. Numbers indicate amino acid positions. (B) Model of IFNγ-FasICD working mechanism. ( C-E ) HEK293T cells were transfected with IFNγ-FasICD mRNA. (C) IFNγ levels in cell lysates were measured by ELISA at 6 h. (D) The expression of membrane-bound IFNγ was analyzed using flow cytometry. (E) IFNγ levels in culture supernatants were measured at 6 h. (F) MC38 and B16OVA cells were transfected with Luc (luciferase), IFNγ-FasICD, or IFNγ-FasΔ mRNA, cell viability was assessed by MTT assay at 24 h. (G-H) B16OVA cells were transfected with the IFNγ-FasICD mRNA. Caspase-3/7 activity was measured to assess apoptosis (G) and Annexin V + cells were quantified (H) . (I) Expression levels of IFNγ receptor 1 and 2 ( Ifngr1 and Ifngr2 ) were analyzed by semi-quantitative RT-PCR. β-actin ( Actb ) was used as an internal control. (J) Expression of IFNγ downstream target genes, Irf1 and Cd274 , was analyzed by real-time PCR. (K) B16OVA cells were transfected with IFNγ-FasICD mRNA and co-cultured with splenocytes from C57BL/6J mice. STAT1 phosphorylation (Tyr701) in splenocytes as analyzed over time by flow cytometry (left and middle panels), and the percentage of pSTAT1 + cells at the 60-min time point among CD45 + cells is shown (right panel). Data are pooled from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C, E, G), ordinary one-way ANOVA followed by Šídák’s multiple comparisons test (J), two-way ANOVA followed by Dunnett’s multiple comparisons test (F), and unpaired t-tests (H, K). Error bars indicate SEM. Note: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: After 24 hours, cell viability was assessed using the MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; GoldBio, T-030-1, USA).

Techniques: Sequencing, Transfection, Enzyme-linked Immunosorbent Assay, Expressing, Membrane, Flow Cytometry, Luciferase, MTT Assay, Activity Assay, Quantitative RT-PCR, Control, Real-time Polymerase Chain Reaction, Cell Culture, Phospho-proteomics