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Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing <t>ORAI1</t> (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.
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Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing <t>ORAI1</t> (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.
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Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing <t>ORAI1</t> (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.
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Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing <t>ORAI1</t> (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.
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Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing <t>ORAI1</t> (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.
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MSC-Exos regulate calcium homeostasis in Tfh. (A) Volcano diagram of DEGs of Tfh between control group and Exo group ( n = 3). (B) KEGG enrichment analysis of Tfh between control group ( n = 3) and Exo group ( n = 3). These pathways were significant differences in the diagram. (C) GO enrichment analysis of DEGs between Tfh and naïve CD4 + T cells in C57 based on the GSE157648 dataset. (D) Fluo-4 AM-loaded Th0 and Tfh treated with/without MSC-Exos in HBSS medium for 30 min, and intracellular Ca 2+ levels were detected by flow cytometry using Fluo-4 AM as the probe ( n = 5). (E) mRNA expression levels of Stim1, Stim2, and <t>Orai1</t> in Tfh with PBS or MSC-Exos coculture were detected by RT-qPCR. (F) WB analysis of Stim1 and Orai1 protein expression in Th0 and Tfh after treatment with MSC-Exos or PBS. (G) WB analysis of Stim1 and Orai1 protein expression in Tfh treated with/without MSC-Exos, and ionomycin (5 μM) was used to elevate intracellular Ca 2+ levels. (H) RT-qPCR analysis of NFATc1, NFATc2, and CaN mRNA expression levels in Tfh after treatment with MSC-Exos or PBS. (I) WB analysis of CaN and NFATc2 protein expression in Tfh treated with/without MSC-Exos. (J) WB analysis of IκB, p-IκB, P65, and p-P65 protein expression in Tfh treated with/without MSC-Exos. Bars indicate the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined by one-way ANOVA with Tukey’s multiple comparisons test.
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MSC-Exos regulate calcium homeostasis in Tfh. (A) Volcano diagram of DEGs of Tfh between control group and Exo group ( n = 3). (B) KEGG enrichment analysis of Tfh between control group ( n = 3) and Exo group ( n = 3). These pathways were significant differences in the diagram. (C) GO enrichment analysis of DEGs between Tfh and naïve CD4 + T cells in C57 based on the GSE157648 dataset. (D) Fluo-4 AM-loaded Th0 and Tfh treated with/without MSC-Exos in HBSS medium for 30 min, and intracellular Ca 2+ levels were detected by flow cytometry using Fluo-4 AM as the probe ( n = 5). (E) mRNA expression levels of Stim1, Stim2, and <t>Orai1</t> in Tfh with PBS or MSC-Exos coculture were detected by RT-qPCR. (F) WB analysis of Stim1 and Orai1 protein expression in Th0 and Tfh after treatment with MSC-Exos or PBS. (G) WB analysis of Stim1 and Orai1 protein expression in Tfh treated with/without MSC-Exos, and ionomycin (5 μM) was used to elevate intracellular Ca 2+ levels. (H) RT-qPCR analysis of NFATc1, NFATc2, and CaN mRNA expression levels in Tfh after treatment with MSC-Exos or PBS. (I) WB analysis of CaN and NFATc2 protein expression in Tfh treated with/without MSC-Exos. (J) WB analysis of IκB, p-IκB, P65, and p-P65 protein expression in Tfh treated with/without MSC-Exos. Bars indicate the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined by one-way ANOVA with Tukey’s multiple comparisons test.
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Image Search Results


Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing ORAI1 (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.

Journal: Scientific Reports

Article Title: Remodeling of calcium signaling and store operated calcium entry as a consequence of human neural progenitor cell differentiation

doi: 10.1038/s41598-025-22675-y

Figure Lengend Snippet: Remodelling of SOCE components in ReNcell VM as a consequence of ReNcell VM cell differentiation. ( a ) Relative expression of ORAI and STIM isoforms in proliferative and 2, 4, 7 and 14-days differentiated ReNcell VM (three independent experiments, *indicates significance in comparison to day 0, one-way ANOVA with Tukey’s multiple comparison’s test, * p < 0.05). ( b ) Representative immunoblotting of proliferative and 14 days differentiated ReNcell VM, showing ORAI1 (top), STIM1 (middle) and β-Actin (bottom) protein expression levels. Uncropped original images are found in Supplementary Fig. . ( c ) Quantification of ORAI1 and STIM1 protein expression relative to β-Actin. Data are mean ± SD of three independent experiments. * indicates significance in comparison to proliferative ReNcell VM using a Student’s t-test, * p < 0.05.

Article Snippet: TaqMan gene expression assays used for this study were GFAP: Hs00909233_m1, MAP2: Hs00258900_m1, ORAI1: Hs03046013_m1, ORAI2: Hs00259863_m1, ORAI3: Hs00743683_s1, STIM1: Hs00162394_m1, STIM2: Hs00372712_m1 (ThermoFisher Scientific).

Techniques: Cell Differentiation, Expressing, Comparison, Western Blot

MSC-Exos regulate calcium homeostasis in Tfh. (A) Volcano diagram of DEGs of Tfh between control group and Exo group ( n = 3). (B) KEGG enrichment analysis of Tfh between control group ( n = 3) and Exo group ( n = 3). These pathways were significant differences in the diagram. (C) GO enrichment analysis of DEGs between Tfh and naïve CD4 + T cells in C57 based on the GSE157648 dataset. (D) Fluo-4 AM-loaded Th0 and Tfh treated with/without MSC-Exos in HBSS medium for 30 min, and intracellular Ca 2+ levels were detected by flow cytometry using Fluo-4 AM as the probe ( n = 5). (E) mRNA expression levels of Stim1, Stim2, and Orai1 in Tfh with PBS or MSC-Exos coculture were detected by RT-qPCR. (F) WB analysis of Stim1 and Orai1 protein expression in Th0 and Tfh after treatment with MSC-Exos or PBS. (G) WB analysis of Stim1 and Orai1 protein expression in Tfh treated with/without MSC-Exos, and ionomycin (5 μM) was used to elevate intracellular Ca 2+ levels. (H) RT-qPCR analysis of NFATc1, NFATc2, and CaN mRNA expression levels in Tfh after treatment with MSC-Exos or PBS. (I) WB analysis of CaN and NFATc2 protein expression in Tfh treated with/without MSC-Exos. (J) WB analysis of IκB, p-IκB, P65, and p-P65 protein expression in Tfh treated with/without MSC-Exos. Bars indicate the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined by one-way ANOVA with Tukey’s multiple comparisons test.

Journal: Biomaterials Research

Article Title: Mesenchymal Stem Cell-Derived Exosomes Inhibit Stim1–Orai1 Signaling and Calcium Overload-Induced Mitochondrial Damage of Follicular Helper T Cells in Lupus

doi: 10.34133/bmr.0255

Figure Lengend Snippet: MSC-Exos regulate calcium homeostasis in Tfh. (A) Volcano diagram of DEGs of Tfh between control group and Exo group ( n = 3). (B) KEGG enrichment analysis of Tfh between control group ( n = 3) and Exo group ( n = 3). These pathways were significant differences in the diagram. (C) GO enrichment analysis of DEGs between Tfh and naïve CD4 + T cells in C57 based on the GSE157648 dataset. (D) Fluo-4 AM-loaded Th0 and Tfh treated with/without MSC-Exos in HBSS medium for 30 min, and intracellular Ca 2+ levels were detected by flow cytometry using Fluo-4 AM as the probe ( n = 5). (E) mRNA expression levels of Stim1, Stim2, and Orai1 in Tfh with PBS or MSC-Exos coculture were detected by RT-qPCR. (F) WB analysis of Stim1 and Orai1 protein expression in Th0 and Tfh after treatment with MSC-Exos or PBS. (G) WB analysis of Stim1 and Orai1 protein expression in Tfh treated with/without MSC-Exos, and ionomycin (5 μM) was used to elevate intracellular Ca 2+ levels. (H) RT-qPCR analysis of NFATc1, NFATc2, and CaN mRNA expression levels in Tfh after treatment with MSC-Exos or PBS. (I) WB analysis of CaN and NFATc2 protein expression in Tfh treated with/without MSC-Exos. (J) WB analysis of IκB, p-IκB, P65, and p-P65 protein expression in Tfh treated with/without MSC-Exos. Bars indicate the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined by one-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: These membranes were blocked with 5% nonfat milk at 25 °C for 1 h. Primary antibodies targeting CaN (Proteintech, 13422-1-AP, China), NFATc2 (Proteintech, 22023-1-AP, China), P65 (Affinity, AF5006, USA), p-P65 (Affinity, AF2006, USA), IκB (Abmart, T55026 , China), p-IκB (Abmart, TA2002, China), Stim1 (Proteintech, 11565-1-AP, China), Orai1 (Proteintech, 28411-1-AP, China), and MCU (Abways, BY0101, China) were incubated overnight at 4 °C.

Techniques: Control, Flow Cytometry, Expressing, Quantitative RT-PCR

MSC-Exos regulate Tfh differentiation and function via a multi-target network, restoring the Tfh/Tfr balance and alleviating SLE. Mechanistically, MSC-Exos inhibit Tfh activation by correcting intracellular calcium dysregulation and preventing mitochondrial calcium overload. In the cytoplasm, MSC-Exos restrict calcium influx through inhibition of the Stim1/Orai1 expression, thereby reducing NFAT and NF-κB activation. In the mitochondria, MSC-Exos suppress aberrant MCU expression, mitigating calcium overload-induced mitochondrial damage, and restoring mitochondrial homeostasis in Tfh. Figure was created with BioGDP .

Journal: Biomaterials Research

Article Title: Mesenchymal Stem Cell-Derived Exosomes Inhibit Stim1–Orai1 Signaling and Calcium Overload-Induced Mitochondrial Damage of Follicular Helper T Cells in Lupus

doi: 10.34133/bmr.0255

Figure Lengend Snippet: MSC-Exos regulate Tfh differentiation and function via a multi-target network, restoring the Tfh/Tfr balance and alleviating SLE. Mechanistically, MSC-Exos inhibit Tfh activation by correcting intracellular calcium dysregulation and preventing mitochondrial calcium overload. In the cytoplasm, MSC-Exos restrict calcium influx through inhibition of the Stim1/Orai1 expression, thereby reducing NFAT and NF-κB activation. In the mitochondria, MSC-Exos suppress aberrant MCU expression, mitigating calcium overload-induced mitochondrial damage, and restoring mitochondrial homeostasis in Tfh. Figure was created with BioGDP .

Article Snippet: These membranes were blocked with 5% nonfat milk at 25 °C for 1 h. Primary antibodies targeting CaN (Proteintech, 13422-1-AP, China), NFATc2 (Proteintech, 22023-1-AP, China), P65 (Affinity, AF5006, USA), p-P65 (Affinity, AF2006, USA), IκB (Abmart, T55026 , China), p-IκB (Abmart, TA2002, China), Stim1 (Proteintech, 11565-1-AP, China), Orai1 (Proteintech, 28411-1-AP, China), and MCU (Abways, BY0101, China) were incubated overnight at 4 °C.

Techniques: Activation Assay, Inhibition, Expressing