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Activity Assay
CCK-8 Assay
Concentration Assay
Electron Microscopy
Enzyme-linked Immunosorbent Assay
Expressing
Immunofluorescence
Immunohistochemistry
Plasmid Preparation
Real-time Polymerase Chain Reaction
Staining
Transfection
Transmission Assay
CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD 
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Investigation of the association between the expression of plasmalogen biosynthesis and ferroptosis in COPD mice. ( A ) H&E staining of sections of mouse lung tissues from COPD model and normal group. ( B ) The concentration of IL-33 and IL-1α in the bronchoalveolar lavage fluid was determined by ELISA assay. ( C ) The neutrophils, macrophages, and lymphocytes in bronchoalveolar lavage fluid were quantified and compared between COPD and normal groups. ( D ) Tissue damage of lung were elevated by immunohistochemistry staingin <t>of</t> <t>Bax.</t> ( E ) GPX4 activity and the contents of GSH and MDA were analyzed between COPD and normal groups. ( F ) GPX4 expression in the lung tissues of mice was tested by immunohistochemistry (×100 and ×200). ( G ) The protein expression of GPX4, FAR-1, AGPS, and <t>GNPAT</t> was detected via western blot analysis. ( H ) The mRNA expression of FAR-1, AGPS, and GNPAT was measured via quantitative real-time PCR. ( I ) GNPAT expression in the lung tissues of mice was tested by immunohistochemistry (×100 and ×200). Data are presented as the mean ± SD of three replicates and analyzed using Student’s t-test. ns p > 0.05, ** p < 0.01, *** p < 0.001, compared with normal. The gels were cropped reasonably
CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD 
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CSE triggered ferroptosis and upregulated the expression of plasmalogen biosynthesis in human alveolar epithelial cells. Human type II alveolar epithelial A549 cells were exposed to 0.1%, 0.5%, 2%, and 5% CSE for 24 h. ( A ) Cell viability was detected by CCK-8 assay. ( B ) LDH release was detected by LDH activity assays. ( C ) The concentration of IL-33 and IL-1α in the cellular supernatant was determined by ELISA assay. ( D ) The levels of total ROS were determined using a 2’, 7’-dichlorofluorescein diacetate (DCFH-DA) kit. ( E-G ) Quantification of the ferroptosis-related markers MDA, GSH, <t>and</t> <t>GPX4.</t> ( H ) The protein levels of GPX4, FAR-1, AGPS, and <t>GNPAT</t> were detected by western blot analysis. ( I ) The mRNA expression of FAR-1, AGPS, and GNPAT was measured via quantitative real-time PCR. Data are presented as the mean ± SD of three replicates and analyzed using one-way analysis of variance (ANOVA), followed by Dunnett’s test. ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, compared with blank. The gels were cropped reasonably
CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD 
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Experimental validation of CSE-induced markers associated with ferroptosis and plasmalogen biosynthesis. ( A ) Mitochondrial morphology associated with ferroptosis was determined by transmission electron microscopy. ( B ) Content detection of the ferroptosis-related markers MDA, GSH, <t>and</t> <t>GPX4</t> by ELISA. ( C ) The mRNA expression of <t>GNPAT</t> was measured via quantitative real-time PCR. ( D ) The protein levels of GPX4 and GNPAT were detected by western blot analysis. ( E ) Immunofluorescence staining of GNPAT was displayed in different groups. Data are presented as the mean ± SD of three replicates and analyzed using ANOVA, followed by Tukey’s post-hoc test. * p < 0.05, *** p < 0.001, compared with blank; $ p < 0.05, compared with 5% CSE; ## p < 0.01, ### p < 0.001, compared with 5% CSE + DMSO. The gels were cropped reasonably
CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD Respiratory Research, 2023 Dec 01
"Investigation of the association between the expression of plasmalogen biosynthesis and ferroptosis in COPD mice. ( A ) H&E staining of sections of mouse lung tissues from COPD model and normal group. ( B ) The concentration of IL-33 and IL-1α in the bronchoalveolar lavage fluid was determined by ELISA assay. ( C ) The neutrophils, macrophages, and lymphocytes in bronchoalveolar lavage fluid were quantified and compared between COPD and normal groups. ( D ) Tissue damage of lung were elevated by immunohistochemistry staingin <t>of</t> <t>Bax.</t> ( E ) GPX4 activity and the contents of GSH and MDA were analyzed between COPD and normal groups. ( F ) GPX4 expression in the lung tissues of mice was tested by immunohistochemistry (×100 and ×200). ( G ) The protein expression of GPX4, FAR-1, AGPS, and <t>GNPAT</t> was detected via western blot analysis. ( H ) The mRNA expression of FAR-1, AGPS, and GNPAT was measured via quantitative real-time PCR. ( I ) GNPAT expression in the lung tissues of mice was tested by immunohistochemistry (×100 and ×200). Data are presented as the mean ± SD of three replicates and analyzed using Student’s t-test. ns p > 0.05, ** p < 0.01, *** p < 0.001, compared with normal. The gels were cropped reasonably "
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