Cleaved caspase 3 asp175 antibody alexa fluor 488 conjugate

Vitamin C (purity: > 99.9%) was purchased from Shandong Xinhua pharmaceutical CO., ltd. DMEM Medium, Fetal Bovine Serum (FBS), Wright-Giemsa staining, and Albumin Bovine V were purchased from MACGENE. RBC lysis Buffer, Sulforhodamine B (SRB), RNase A, Propidium Iodide (PI), and rat IgG were purchased from Solarbio. Triton X-100 solution, Ca²⁺ specific fluorescent probe Fluo-4/AM, JC-1 Staining Kit, One Step TUNEL Apoptosis Assay Kit, and ATP Determination Kit were purchased from Beyotime. FITC Rat Anti-Mouse CD11b was purchased from BD Biosciences. APC Rat Anti-Mouse CD206 was purchased from Miltenyi Biotec. Western blot Antibody Diluent was purchased from Epizyme. Western Bright™ ECL and Western Bright™ Peroxide were purchased from Advansta. Goat Anti-Rabbit IgG (H+L) HRP was purchased from Affinity Biosciences. Vimentin (5G3F10) Mouse mAb, Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor^® 488 Conjugate) and β-actin (13E5) Rabbit mAb were purchased from Cell Signaling Technology. E-cadherin Rabbit PolyAb was purchased from Proteintech.

JC-1(5, 50, 6, 60-tetrachloro-1, 10, 3, 30-tetraethylbenzamidazolocarbocyanin iodide, Beyotime Institute Biotechnology) was used to evaluate the change in mitochondrial membrane potential as described previously [24 ]. A549 cells in the logarithmic growth phase were plated in 6-well cell culture plates and incubated for 24 hours, and then transfected with negative control or GCLC-specific siRNA-1. The transfected cells were treated with or without GNPs (20μM) for 72 hours, subsequently the cells were harvested and incubated with JC-1 for 30 minutes in darkness at 37°C. After staining, cells were washed twice with PBS and analysed by flow cytometry [34 (link)].
For intracellular cleaved caspase-3, the transfected cells that treated with or without GNPs were collected and fixed with 4% paraformaldehyde. After treated with 0.1% Triton X-100 and blocked with 1% BSA, cells were incubated with cleaved caspase-3 (Asp175) antibody (Alexa fluor 488 conjugate, Cell Signaling Technology) for 30 minutes. Then, the stained cells were analysed by flow cytometry [24 ].