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calcein am pi double staining solution  (Beyotime)


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    Beyotime calcein am pi double staining solution
    Oba suppresses microglial inflammation and blocks the subsequent apoptosis of HT22 neurons. ( A ) Viability of BV-2 cells treated with different concentrations of Oba, as assessed by CCK-8 assay (n = 5). ( B ) Representative <t>Calcein</t> AM (green)/PI (red) staining images of BV-2 cells following treatment with Oba at the indicated concentrations (0, 25, and 50 μM). Scale bar = 100 μm. ( C ) Quantitative analysis of the ratio of viable cells (n = 5). ( D ) Representative Western blot images of iNOS, TNFα, and IL-1β expression in BV-2 cells. ( E ) Quantitative analysis of iNOS, TNFα and IL-1β protein levels (n = 5). ( F ) Representative immunofluorescence images of iNOS (green) and DAPI (blue) in BV-2 cells treated as follows: vehicle (Control), LPS, LPS+Oba 25 μM, and LPS+Oba 50 μM. Scale bar = 50 μm. ( G ) Quantitative analysis of fluorescence intensity of iNOS (n = 5). ( H ) mRNA expression levels of TNFα and IL-1β in BV-2 cells measured by qPCR (n = 6). ( I ) Representative Western blot image of Bax and Cleaved-caspase3 protein level in HT22 cells. ( J ) Quantitative analysis of Bax and Cleaved-caspase3 protein levels (n = 5). ( K ) Representative flow cytometry plots of HT22 cells stained with Annexin V-FITC (x-axis) and PI (y-axis). ( L ) The percentage of apoptotic HT22 cells, as determined by flow cytometry. (n = 5). The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, 25 μM Oba, and 50 μM Oba. The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.
    Calcein Am Pi Double Staining Solution, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 3029 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/calcein+am+staining+solution/Calcein+AM/pmc12994538-118-5-11
    Average 99 stars, based on 3029 article reviews
    calcein am pi double staining solution - by Bioz Stars, 2026-09
    99/100 stars

    Images

    1) Product Images from "Obacunone Promotes Functional Recovery After Spinal Cord Injury by Attenuating Neuroinflammation by Targeting the TLR4/MyD88/p38 MAPK Pathway"

    Article Title: Obacunone Promotes Functional Recovery After Spinal Cord Injury by Attenuating Neuroinflammation by Targeting the TLR4/MyD88/p38 MAPK Pathway

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S577707

    Oba suppresses microglial inflammation and blocks the subsequent apoptosis of HT22 neurons. ( A ) Viability of BV-2 cells treated with different concentrations of Oba, as assessed by CCK-8 assay (n = 5). ( B ) Representative Calcein AM (green)/PI (red) staining images of BV-2 cells following treatment with Oba at the indicated concentrations (0, 25, and 50 μM). Scale bar = 100 μm. ( C ) Quantitative analysis of the ratio of viable cells (n = 5). ( D ) Representative Western blot images of iNOS, TNFα, and IL-1β expression in BV-2 cells. ( E ) Quantitative analysis of iNOS, TNFα and IL-1β protein levels (n = 5). ( F ) Representative immunofluorescence images of iNOS (green) and DAPI (blue) in BV-2 cells treated as follows: vehicle (Control), LPS, LPS+Oba 25 μM, and LPS+Oba 50 μM. Scale bar = 50 μm. ( G ) Quantitative analysis of fluorescence intensity of iNOS (n = 5). ( H ) mRNA expression levels of TNFα and IL-1β in BV-2 cells measured by qPCR (n = 6). ( I ) Representative Western blot image of Bax and Cleaved-caspase3 protein level in HT22 cells. ( J ) Quantitative analysis of Bax and Cleaved-caspase3 protein levels (n = 5). ( K ) Representative flow cytometry plots of HT22 cells stained with Annexin V-FITC (x-axis) and PI (y-axis). ( L ) The percentage of apoptotic HT22 cells, as determined by flow cytometry. (n = 5). The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, 25 μM Oba, and 50 μM Oba. The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.
    Figure Legend Snippet: Oba suppresses microglial inflammation and blocks the subsequent apoptosis of HT22 neurons. ( A ) Viability of BV-2 cells treated with different concentrations of Oba, as assessed by CCK-8 assay (n = 5). ( B ) Representative Calcein AM (green)/PI (red) staining images of BV-2 cells following treatment with Oba at the indicated concentrations (0, 25, and 50 μM). Scale bar = 100 μm. ( C ) Quantitative analysis of the ratio of viable cells (n = 5). ( D ) Representative Western blot images of iNOS, TNFα, and IL-1β expression in BV-2 cells. ( E ) Quantitative analysis of iNOS, TNFα and IL-1β protein levels (n = 5). ( F ) Representative immunofluorescence images of iNOS (green) and DAPI (blue) in BV-2 cells treated as follows: vehicle (Control), LPS, LPS+Oba 25 μM, and LPS+Oba 50 μM. Scale bar = 50 μm. ( G ) Quantitative analysis of fluorescence intensity of iNOS (n = 5). ( H ) mRNA expression levels of TNFα and IL-1β in BV-2 cells measured by qPCR (n = 6). ( I ) Representative Western blot image of Bax and Cleaved-caspase3 protein level in HT22 cells. ( J ) Quantitative analysis of Bax and Cleaved-caspase3 protein levels (n = 5). ( K ) Representative flow cytometry plots of HT22 cells stained with Annexin V-FITC (x-axis) and PI (y-axis). ( L ) The percentage of apoptotic HT22 cells, as determined by flow cytometry. (n = 5). The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, 25 μM Oba, and 50 μM Oba. The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.

    Techniques Used: CCK-8 Assay, Staining, Western Blot, Expressing, Immunofluorescence, Control, Fluorescence, Flow Cytometry

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    Incubation:

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    Article Title: A Fluorescent Probe for Imaging and Treating S-Nitrosation Stress in OGD/R Cells
    Article Snippet: .. Subsequently, Calcein AM staining solution (final concentration 2 μM) (Beyotime, Shanghai, China) was added to the culture medium and incubated for 15 min. Propidium iodide (PI, final concentration 50 μg/mL) (Biofroxx, Einhausen, Germany) was then added for a 5 min incubation period. ..

    Article Title: A Fluorescent Probe for Imaging and Treating S-Nitrosation Stress in OGD/R Cells
    Article Snippet: .. Subsequently, Calcein AM staining solution (final concentration 2 μM) (Beyotime, Shanghai, China) was added to the culture medium and incubated for 15 min. Propidium iodide (PI, final concentration 50 μg/mL) (Biofroxx, Einhausen, Germany) was then added for a 5 min incubation period. ..

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    Article Title: VDAC1 Oligomerization-Mediated mtDNA Release under Sublethal Oxidative Stress: A Novel Inflammatory Mechanism in Vitiligo.
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    Cell Culture:

    Article Title: An injectable hydrogel loaded with miRNA nanocarriers promotes vessel-associated osteoclast (VAO)-mediated angiogenesis and bone regeneration in osteonecrosis of the rat femoral head.
    Article Snippet: Osteonecrosis of the femoral head (ONFH) remains a significant clinical challenge.. Despite various strategies aimed at promoting bone repair and halting disease progression, an effective cure remains elusive.. Recent studies have identified a non-bone-resorbing osteoclast subtype, vessel-associated osteoclasts (VAOs), distinct from classical bone-associated osteoclasts (BAOs), offering new therapeutic opportunities for ONFH.

    Staining:

    Article Title: An injectable hydrogel loaded with miRNA nanocarriers promotes vessel-associated osteoclast (VAO)-mediated angiogenesis and bone regeneration in osteonecrosis of the rat femoral head.
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    Article Snippet: .. Cells were treated using DCFH-DA (Beyotime, China), Calcein AM staining solution (Beyotime, China), MitosoxTMRed (ThermoFisher, USA), and mitochondrial membrane potential (Solarbio, China) according to the manufacturer's procedure before image capture using the microscope. .. Protein samples that have been prepared were run on precast SDS-PAGE gels (Vazyme, China) and electrophoretically converted onto the polyvinylidene difluoride membrane.

    Article Title: A Fluorescent Probe for Imaging and Treating S-Nitrosation Stress in OGD/R Cells
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    Article Title: A Fluorescent Probe for Imaging and Treating S-Nitrosation Stress in OGD/R Cells
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    Article Title: Icaritin alleviates UVB-induced skin damage by inhibiting ferroptosis via modulation of mitochondrial dynamics.
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    Fluorescence:

    Article Title: An injectable hydrogel loaded with miRNA nanocarriers promotes vessel-associated osteoclast (VAO)-mediated angiogenesis and bone regeneration in osteonecrosis of the rat femoral head.
    Article Snippet: Osteonecrosis of the femoral head (ONFH) remains a significant clinical challenge.. Despite various strategies aimed at promoting bone repair and halting disease progression, an effective cure remains elusive.. Recent studies have identified a non-bone-resorbing osteoclast subtype, vessel-associated osteoclasts (VAOs), distinct from classical bone-associated osteoclasts (BAOs), offering new therapeutic opportunities for ONFH.

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    Microscopy:

    Article Title: An injectable hydrogel loaded with miRNA nanocarriers promotes vessel-associated osteoclast (VAO)-mediated angiogenesis and bone regeneration in osteonecrosis of the rat femoral head.
    Article Snippet: Osteonecrosis of the femoral head (ONFH) remains a significant clinical challenge.. Despite various strategies aimed at promoting bone repair and halting disease progression, an effective cure remains elusive.. Recent studies have identified a non-bone-resorbing osteoclast subtype, vessel-associated osteoclasts (VAOs), distinct from classical bone-associated osteoclasts (BAOs), offering new therapeutic opportunities for ONFH.

    Article Title: Rg1-R1 attenuates cardiac ischemia/reperfusion-induced endothelial cell injury through activating the ULK1/PGAM5-FUNDC1-mitophagy pathway
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    Membrane:

    Article Title: Rg1-R1 attenuates cardiac ischemia/reperfusion-induced endothelial cell injury through activating the ULK1/PGAM5-FUNDC1-mitophagy pathway
    Article Snippet: .. Cells were treated using DCFH-DA (Beyotime, China), Calcein AM staining solution (Beyotime, China), MitosoxTMRed (ThermoFisher, USA), and mitochondrial membrane potential (Solarbio, China) according to the manufacturer's procedure before image capture using the microscope. .. Protein samples that have been prepared were run on precast SDS-PAGE gels (Vazyme, China) and electrophoretically converted onto the polyvinylidene difluoride membrane.

    Concentration Assay:

    Article Title: A Fluorescent Probe for Imaging and Treating S-Nitrosation Stress in OGD/R Cells
    Article Snippet: .. Subsequently, Calcein AM staining solution (final concentration 2 μM) (Beyotime, Shanghai, China) was added to the culture medium and incubated for 15 min. Propidium iodide (PI, final concentration 50 μg/mL) (Biofroxx, Einhausen, Germany) was then added for a 5 min incubation period. ..

    Article Title: A Fluorescent Probe for Imaging and Treating S-Nitrosation Stress in OGD/R Cells
    Article Snippet: .. Subsequently, Calcein AM staining solution (final concentration 2 μM) (Beyotime, Shanghai, China) was added to the culture medium and incubated for 15 min. Propidium iodide (PI, final concentration 50 μg/mL) (Biofroxx, Einhausen, Germany) was then added for a 5 min incubation period. ..

    ROS Assay:

    Article Title: Integrated UPLC-ESI-MS/MS, network pharmacology, and transcriptomics to reveal the material basis and mechanism of Schisandra chinensis Fruit Mixture against diabetic nephropathy
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    Elabscience Biotechnology vortex calcein am solution
    Rg1-R1 alleviates H/R-induced endothelial <t>cell</t> <t>mitochondrial</t> dysfunction by culturing and isolating CMECs. (A) Changes in mitochondria and autophagic vesicles of endothelial cells in each group were observed using transmission electron microscopy. Red arrows indicate mitochondria and yellow arrows indicate mitochondria wrapped by autophagic lysosomes. (B) OCR measurement was used to observe cellular respiratory function and the production of ATP with or without Rg1-R1 treatment. (C) Mitochondria ROS were gauged by Mitosox™Red and cytoplasmic ROS were gauged by H2DCF-DA. (D) JC-1 was utilised to monitor the membrane potential of the mitochondria. (E) The openness of mPTP was determined based on the intensity of green fluorescence of <t>calcein</t> acetoxymethyl esterin mitochondria. (F) Expression of p-Drp1, Mff, Fis1, Mfn2, and Opa1 was examined by Western blot. Experiments were repeated at least three times and data are shown asmean ± SD (Three independent cellular alleles per group). ∗∗p < 0.01, ∗p < 0.05.
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    Oba suppresses microglial inflammation and blocks the subsequent apoptosis of HT22 neurons. ( A ) Viability of BV-2 cells treated with different concentrations of Oba, as assessed by CCK-8 assay (n = 5). ( B ) Representative Calcein AM (green)/PI (red) staining images of BV-2 cells following treatment with Oba at the indicated concentrations (0, 25, and 50 μM). Scale bar = 100 μm. ( C ) Quantitative analysis of the ratio of viable cells (n = 5). ( D ) Representative Western blot images of iNOS, TNFα, and IL-1β expression in BV-2 cells. ( E ) Quantitative analysis of iNOS, TNFα and IL-1β protein levels (n = 5). ( F ) Representative immunofluorescence images of iNOS (green) and DAPI (blue) in BV-2 cells treated as follows: vehicle (Control), LPS, LPS+Oba 25 μM, and LPS+Oba 50 μM. Scale bar = 50 μm. ( G ) Quantitative analysis of fluorescence intensity of iNOS (n = 5). ( H ) mRNA expression levels of TNFα and IL-1β in BV-2 cells measured by qPCR (n = 6). ( I ) Representative Western blot image of Bax and Cleaved-caspase3 protein level in HT22 cells. ( J ) Quantitative analysis of Bax and Cleaved-caspase3 protein levels (n = 5). ( K ) Representative flow cytometry plots of HT22 cells stained with Annexin V-FITC (x-axis) and PI (y-axis). ( L ) The percentage of apoptotic HT22 cells, as determined by flow cytometry. (n = 5). The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, 25 μM Oba, and 50 μM Oba. The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.

    Journal: Drug Design, Development and Therapy

    Article Title: Obacunone Promotes Functional Recovery After Spinal Cord Injury by Attenuating Neuroinflammation by Targeting the TLR4/MyD88/p38 MAPK Pathway

    doi: 10.2147/DDDT.S577707

    Figure Lengend Snippet: Oba suppresses microglial inflammation and blocks the subsequent apoptosis of HT22 neurons. ( A ) Viability of BV-2 cells treated with different concentrations of Oba, as assessed by CCK-8 assay (n = 5). ( B ) Representative Calcein AM (green)/PI (red) staining images of BV-2 cells following treatment with Oba at the indicated concentrations (0, 25, and 50 μM). Scale bar = 100 μm. ( C ) Quantitative analysis of the ratio of viable cells (n = 5). ( D ) Representative Western blot images of iNOS, TNFα, and IL-1β expression in BV-2 cells. ( E ) Quantitative analysis of iNOS, TNFα and IL-1β protein levels (n = 5). ( F ) Representative immunofluorescence images of iNOS (green) and DAPI (blue) in BV-2 cells treated as follows: vehicle (Control), LPS, LPS+Oba 25 μM, and LPS+Oba 50 μM. Scale bar = 50 μm. ( G ) Quantitative analysis of fluorescence intensity of iNOS (n = 5). ( H ) mRNA expression levels of TNFα and IL-1β in BV-2 cells measured by qPCR (n = 6). ( I ) Representative Western blot image of Bax and Cleaved-caspase3 protein level in HT22 cells. ( J ) Quantitative analysis of Bax and Cleaved-caspase3 protein levels (n = 5). ( K ) Representative flow cytometry plots of HT22 cells stained with Annexin V-FITC (x-axis) and PI (y-axis). ( L ) The percentage of apoptotic HT22 cells, as determined by flow cytometry. (n = 5). The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, 25 μM Oba, and 50 μM Oba. The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.

    Article Snippet: Subsequently, an appropriate volume of Calcein AM/PI double staining solution (C2015S, Beyotime, China) was added to each well.

    Techniques: CCK-8 Assay, Staining, Western Blot, Expressing, Immunofluorescence, Control, Fluorescence, Flow Cytometry

    Rg1-R1 alleviates H/R-induced endothelial cell mitochondrial dysfunction by culturing and isolating CMECs. (A) Changes in mitochondria and autophagic vesicles of endothelial cells in each group were observed using transmission electron microscopy. Red arrows indicate mitochondria and yellow arrows indicate mitochondria wrapped by autophagic lysosomes. (B) OCR measurement was used to observe cellular respiratory function and the production of ATP with or without Rg1-R1 treatment. (C) Mitochondria ROS were gauged by Mitosox™Red and cytoplasmic ROS were gauged by H2DCF-DA. (D) JC-1 was utilised to monitor the membrane potential of the mitochondria. (E) The openness of mPTP was determined based on the intensity of green fluorescence of calcein acetoxymethyl esterin mitochondria. (F) Expression of p-Drp1, Mff, Fis1, Mfn2, and Opa1 was examined by Western blot. Experiments were repeated at least three times and data are shown asmean ± SD (Three independent cellular alleles per group). ∗∗p < 0.01, ∗p < 0.05.

    Journal: Journal of Ginseng Research

    Article Title: Rg1-R1 attenuates cardiac ischemia/reperfusion-induced endothelial cell injury through activating the ULK1/PGAM5-FUNDC1-mitophagy pathway

    doi: 10.1016/j.jgr.2025.100973

    Figure Lengend Snippet: Rg1-R1 alleviates H/R-induced endothelial cell mitochondrial dysfunction by culturing and isolating CMECs. (A) Changes in mitochondria and autophagic vesicles of endothelial cells in each group were observed using transmission electron microscopy. Red arrows indicate mitochondria and yellow arrows indicate mitochondria wrapped by autophagic lysosomes. (B) OCR measurement was used to observe cellular respiratory function and the production of ATP with or without Rg1-R1 treatment. (C) Mitochondria ROS were gauged by Mitosox™Red and cytoplasmic ROS were gauged by H2DCF-DA. (D) JC-1 was utilised to monitor the membrane potential of the mitochondria. (E) The openness of mPTP was determined based on the intensity of green fluorescence of calcein acetoxymethyl esterin mitochondria. (F) Expression of p-Drp1, Mff, Fis1, Mfn2, and Opa1 was examined by Western blot. Experiments were repeated at least three times and data are shown asmean ± SD (Three independent cellular alleles per group). ∗∗p < 0.01, ∗p < 0.05.

    Article Snippet: Cells were treated using DCFH-DA (Beyotime, China), Calcein AM staining solution (Beyotime, China), MitosoxTMRed (ThermoFisher, USA), and mitochondrial membrane potential (Solarbio, China) according to the manufacturer's procedure before image capture using the microscope.

    Techniques: Transmission Assay, Electron Microscopy, Membrane, Fluorescence, Expressing, Western Blot