primary antibodies against kidney injury molecule-1 (kim-1)  (Novus Biologicals)

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

Journal: Nutrients

Article Title: Beneficial Effects of Isoflavones in the Kidney of Obese Rats Are Mediated by PPAR-Gamma Expression

doi: 10.3390/nu12061624

Figure Lengend Snippet: Evaluation of renal function. Rats in the CTL group, the DH group, and the DH and ISO group at baseline, 60, 90, and/or 120 days. ( A ) Creatinine clearance, ( B ) plasma urea, ( C ) plasma creatinine, ( D ) uric acid, ( E ) fractional sodium excretion, and ( F ) urinary protein. ( G ) Light microscopy of kidney injury molecule-1 (KIM-1) and quantitative analyses of kidney sections stained for KIM-1. Data are reported as mean ± standard error. The significance level for a null hypothesis was set at 5% ( p < 0.05). (*) compared to the CTL group at 60 days, (**) compared to the DH group at 60 days, (¥) compared to the CTL group at 90 days, (¥¥) compared to the DH group at 90 days, (₸) compared to the CTL group at 120 days, and (₸₸) compared to the DH group at 120 days.

Article Snippet: Paraffin sections were subjected to alcohol and xylene gradient solutions, antigen retrieval, protein block, and incubation with primary antibodies against CD31 (1:200, mouse clone JC70a endothelial, M082, Dako, CA, USA), CD68 (1:200, mouse monoclonal clone, M0876, Dako, CA, USA), and kidney injury molecule 1 (KIM-1) (1:200, rabbit IgG, H07H, Sino Biologica, BJ, CH) overnight at 4 °C.

Techniques: Clinical Proteomics, Light Microscopy, Staining