Anti cd95

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-CD95 is a monoclonal antibody that binds to the CD95 (Fas/Apo-1) receptor. CD95 is a cell surface receptor that plays a key role in the regulation of programmed cell death (apoptosis).

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3 protocols using anti cd95

Comprehensive Antibody Panel for Cell Signaling Analysis

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Catalog numbers and dilution fold are listed in parentheses. The following antibodies were purchased from Cell Signaling Technology: Anti-Bak (3792; 1:1000), anti-Bax (2772; 1:1000), anti-Bcl-2 (2870; 1:1000), anti-Bcl-xL (2764; 1:500), anti-CHK1 (2360; 1:1000), anti-phospho-CHK1 (S345; 2348; 1:1000), anti-cleaved caspase-3 (9661; 1:1000), anti-cleaved caspase-7 (9491; 1:1000), anti-cleaved caspase-8 (9496; 1:500), anti-cleaved caspase-9 (9501; 1:1000), anti-cytochrome c (4272; 1:500), anti-H2A.X (7631; 1:1000), anti-phospho-H2A.X (S139; 9718; 1:1000), anti-p53 (2524; 1:1000), anti-p53 (2527; 1:1000), anti-phospho-p53 (S6; 9285; 1:1000), anti-phospho-p53 (S15; 9284; 1:1000), anti-p21 (2946; 1:1000), and anti-voltage-dependent anion channel (VDAC; 4866; 1:1000). The following antibodies were purchased from Santa Cruz Biotechnology: Anti-CD95 (1023; 1:500), anti-cyclophilin 40, also known as anti-CYPD (137157; 1:400), anti-His tag (803; 1:500), anti-lamin A (20680; 1:1000), anti-MDM2 (965; 1:500), and anti-ubiquitin (8017; 1:200). Anti-PEPD (Ab86507; 1:500) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; MAB374; 1:5000) were purchased from Abcam and EMD Millipore, respectively. Anti-mouse IgG-horseradish peroxidase (IgG-HRP; NA931V; 1:4000–5000) and anti-rabbit IgG-HRP (NA934V; 1:5000) were purchased from GE Healthcare.

Yang L., Li Y., Bhattacharya A, & Zhang Y. (2017). PEPD is a pivotal regulator of p53 tumor suppressor. Nature Communications, 8, 2052.

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Comparative Protein Expression Analysis in Diabetic Rat Tissues

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Hepatic and pancreatic tissue lysates from control and STZ-induced male and female rats containing equal amounts of protein were separated by SDS-PAGE. After electrophoresis, proteins were transferred to polyvinylidene difluoride (PVDF) membranes, blocked for 1h with 5% skim milk at room temperature, and incubated with the indicated primary antibodies for 2 h at 1:1000 dilution (anti-β-actin, anti-SPARC, anti-C/EBPβ, anti-ATP5B, anti-CD95, anti-MCP1, anti-iNOS, anti-Cyt C, anti-SOD2, anti-INS, anti-CA3, anti-RARhoGAP, anti-CPS1, anti-BHMT, anti-PA, anti-APC2 [Santa Cruz Biotechnology, Santa Cruz, CA, USA], anti-TNFα, anti-PARP-1, anti-NFκB, anti-HSP90 [Cell Signaling Technology, Beverly, MA, USA], and anti-CRP [AbFrontier, Seoul, Korea]). After washing with Tris-buffered saline containing Tween 20, the membranes were incubated with HRP-conjugated secondary antibodies for 1 h at room temperature. Then, immune complexes were detected using the ECL method, and immunoreactive bands were quantified by densitometric analysis using ImageMaster 2D software version 4.95 (GE Healthcare, Little Chalfont, Buckinghamshire, UK). Relative intensity (%) values of proteins were normalized to β-actin levels.

Aseer K.R., Kim S.W., Choi M.S, & Yun J.W. (2015). Opposite Expression of SPARC between the Liver and Pancreas in Streptozotocin-Induced Diabetic Rats. PLoS ONE, 10(6), e0131189.

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Serum Starvation and FasL-Induced Apoptosis in MSCs

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MSCs were serum starved for 24 hours, before 0.2 ng/ml FasL was added. Cells were harvested at the indicated time points. To analyse the effects of tumour cell supernatants on MSCs, stem cells were seeded, and 24 hours later, the growth medium was replaced with cell supernatant for the indicated time-points. For caspase western blots hMSCs were treated with 5 ng/ml FasL or 5 ng/ml multimeric-FasL in regular growth medium for 24 hours. SDS-PAGE and transfer onto PVDF membranes were performed as described previously [57] . The following primary antibodies were used: anti-phospho-SAPK/JNK (Cell Signaling Technology, Danvers, MA, USA), anti-total-SAPK/JNK (Cell Signaling Technology), anti-phospho-p38 MAPK (Cell Signaling Technology), anti-total-p38 MAPK (Cell Signaling Technology), anti-phospho-ERK (Cell Signaling Technology), anti-total-ERK (Cell Signaling Technology), anti-CD95 (Santa Cruz Biotechnology, Dallas, TX, USA), anti-IκBα (Cell Signaling Technology), anti-caspase-3 (Biotechne), anti-caspase-8 (Cell Signaling Technology), anti-vimentin (BioLegend) and anti-CuZnSOD (Binding Site, Birmingham, UK). Peroxidase-conjugated secondary antibodies were anti-rabbit and anti-goat sourced from Santa Cruz. All antibodies were diluted to a working concentration of 1:1000. Proteins were visualised using ECL and a Fusion FX Chemidoc Imager (Vilber, Collégien, France).

Mohr A., Chu T., Clarkson C.T., Brooke G.N., Teif V.B, & Zwacka R.M. (2021). Fas-threshold signalling in MSCs promotes pancreatic cancer progression and metastasis. Cancer letters, 519.

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