primary antibodies against kidney injury molecule 1  (Proteintech)

Journal: Renal Failure

Article Title: Magnesium hydride protects against acetaminophen-induced acute kidney injury by inhibiting TXNIP/NLRP3/nf-κb pathway

doi: 10.1080/0886022X.2024.2330629

Figure Lengend Snippet: MgH 2 ameliorates APAP-induced renal dysfunction, histological injury and oxidative stress in mice. (A, B) The levels of SCr and BUN in serum of mice were determined by corresponding kits. (C–E) The renal histological changes in mice were observed by HE staining (C), Masson staining (D) and PAS staining (E). (F, G) The protein expressions of NGAL, KIM-1 and iNOS in kidneys of mice were detected by western blotting. (H) The intracellular ROS level in renal tissues of mice was detected using DHE. The results were expressed as mean ± SEM. Statistical comparisons were performed using a Newman–Keuls test (* p < 0.05 vs. Control group, # p < 0.05 vs. APAP group).

Article Snippet: After blocking in 5% bovine serum for 2 h, the membranes were incubated at 4 °C overnight with primary antibodies against neutrophil gelatinase-associated lipocalin (NGAL; 1:1000, Cloud-Clone, China), kidney injury molecule-1 (KIM-1; 1:1000, Cloud-Clone, China), inducible nitric oxide synthase (iNOS; 1:1000, Cell Signaling Technology, USA), tumor necrosis factor-α (TNF-α; 1:1000, Servicebio, China), interleukin-1β (IL-1β; 1:1000, Cloud-Clone, China), Bcl2-antagonist of cell death (Bad; 1:1000, Cloud-Clone, China), Bcl2 associated X protein (Bax; 1:1000, Cell Signaling Technology, USA), Caspase3 (1:1000, Proteintech, USA), cytochrome C (CytC; 1:1000, Cell Signaling Technology, USA), TXNIP (1:1000), NLRP3 (1:1000, Cell Signaling Technology, USA), NF-κB p65 (1:1000, Cell Signaling Technology, USA), p-NF-κB p65 (1:1000), β-Actin (1:1000, Cell Signaling Technology, USA), β-Tubulin (1:1000, Abways, China) and GAPDH (1:1000, Servicebio, China).

Techniques: Staining, Western Blot

Journal: Renal Failure

Article Title: Magnesium hydride protects against acetaminophen-induced acute kidney injury by inhibiting TXNIP/NLRP3/nf-κb pathway

doi: 10.1080/0886022X.2024.2330629

Figure Lengend Snippet: MgH 2 alleviates APAP-induced cytotoxicity, oxidative stress, mitochondrial dysfunction and inflammation in HK-2 cells. (A-C) HK-2 Cell viability was detected by CCK-8 assay. (A) HK-2 cells was administered with different concentrations of MgH 2 (10, 20, 50, 100, 200, 400, 600, 800 and 1000 μM); (B) HK-2 cells was administered with different concentrations of APAP (5, 10, 20, 40, 60, 80, 100 mM) with or without MgH 2 (0.4 mM); (C) HK-2 cells was administered with APAP (10 mM) with or without MgH 2 (0.2, 0.4 mM). (D) The ROS level in HK-2 cells was determined by DCFH-DA. (E) The mitochondrial function of HK-2 cells was assessed by JC-1 staining. (F, G) The protein expressions of iNOS, KIM-1 and IL-1β in HK-2 cells were detected by western blotting. The results were expressed as mean ± SEM. Statistical comparisons were performed using t -test (* p < 0.05 vs. APAP group) or Newman–Keuls test (* p < 0.05 vs. Control group, # p < 0.05 vs. APAP group).

Article Snippet: After blocking in 5% bovine serum for 2 h, the membranes were incubated at 4 °C overnight with primary antibodies against neutrophil gelatinase-associated lipocalin (NGAL; 1:1000, Cloud-Clone, China), kidney injury molecule-1 (KIM-1; 1:1000, Cloud-Clone, China), inducible nitric oxide synthase (iNOS; 1:1000, Cell Signaling Technology, USA), tumor necrosis factor-α (TNF-α; 1:1000, Servicebio, China), interleukin-1β (IL-1β; 1:1000, Cloud-Clone, China), Bcl2-antagonist of cell death (Bad; 1:1000, Cloud-Clone, China), Bcl2 associated X protein (Bax; 1:1000, Cell Signaling Technology, USA), Caspase3 (1:1000, Proteintech, USA), cytochrome C (CytC; 1:1000, Cell Signaling Technology, USA), TXNIP (1:1000), NLRP3 (1:1000, Cell Signaling Technology, USA), NF-κB p65 (1:1000, Cell Signaling Technology, USA), p-NF-κB p65 (1:1000), β-Actin (1:1000, Cell Signaling Technology, USA), β-Tubulin (1:1000, Abways, China) and GAPDH (1:1000, Servicebio, China).

Techniques: CCK-8 Assay, Staining, Western Blot