Human/Mouse FLIP Antibody

Detection of Human/Mouse FLIP by Western Blot.

Western blot shows lysates of WS-1 human fetal skin fibroblast cell line and L-929 mouse fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse FLIP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF821) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for FLIP at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Mouse FLIP by Western Blot

TNF alpha and CHX trigger an apoptotic response in cFlip knockdown 17EM15 cells and ex vivo cultured mouse lens capsular bag by activating caspase-8, caspase-9, and caspase-3.A cFlip knocking down in 17EM15 mouse LECs determined by immunoblot-assay. B The semi-quantitative densitometric measurement of 17EM15 cFlip knocking down from immunoblot-assay. C cFlip knocking down in ex vivo cultured mouse lens capsular bags. D The semi-quantitative densitometric measurement of cFlip knocking down mouse lens capsular bags from immunoblot-assay. E Pro-caspase-8, cleaved caspase-8, pro-caspase-3, and cleaved caspase-3 in cFlip knockdown and scrambled shRNA control (SC) 17EM15 cells with and without 30 ng/ml TNF alpha and 10μg/ml CHX stimulation for 7 h. F Pro-caspase-8, cleaved caspase-8, pro-caspase-3, and cleaved caspase-3 in cFlip knockdown and scrambled shRNA control (SC) ex vivo cultured mouse lens capsular bags with and without 60 ng/ml TNF alpha and 10 μg/ml CHX treatment for 24 h. The cFlip KD 17EM15 treated cell lysate was used as a positive control. Only 1/10th amount of protein relative to capsular bag lysate was loaded. One-way ANOVA was used to compare between groups, and only p < 0.05 is considered significant. *<0.05, **<0.01, ***<0.001, ****<0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33837174), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FLIP by Western Blot

cFLIP is highly expressed in FHL124 cells and also significantly upregulated after TNF alpha stimulation.A Anti-apoptotic protein expression in FHL124 and HeLa cell with and without TNF alpha and TNF alpha plus CHX stimulation. B Relative cFLIP mRNA expression in FHL124 and HeLa cells with and without TNF alpha and CHX stimulation. FHL 124 cells demonstrated a more robust response to TNF alpha and CHX stimulation compared to HeLa cells. C The endogenous cFLIP, Caspase-8, caspase-9, and caspase-3 mRNA levels in FHL124 cells vs. HeLa cells. A much higher cFLIP mRNA expression was seen in FHL124 cells compared to HeLa cells. D,E cFLIP protein expression after TNF alpha and CHX stimulation in FHL124 and HeLa cells. cFLIP protein level was increased around 3-fold in FHL124 cells at 5 h after TNF alpha and CHX stimulation, while only a mild increase was seen in HeLa cells after stimulation. F,G FHL124 cells had 6-fold endogenous cFLIP protein levels vs. HeLa cells. KD3 shRNA was able to knock down 50% of cFLIP expression in FHL124 cells. Each assay was repeated at least three times. One-way ANOVA with Tukey’s Honest post-hoc analysis was used to compare between groups, and only p < 0.05 is considered significant. *<0.05, **<0.01, ***<0.001, ****<0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33837174), licensed under a CC-BY license. Not internally tested by R&D Systems.