Pe conjugated anti fas

Manufactured by BD

Sourced in United States

The PE-conjugated anti-Fas is a fluorescently-labeled monoclonal antibody that binds to the Fas receptor (CD95) on the cell surface. It can be used to detect and quantify Fas expression on various cell types in flow cytometry applications.

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4 protocols using pe conjugated anti fas

Sirt3 Regulates Germinal Center Formation

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Control Sirt3^+/+ and Sirt3^−/− mice were immunized intraperitoneally at 8 to 12 weeks of age with 0.5 ml suspension of 2% sheep red blood cell (SRBC) in PBS (Cocalico Biologicals) to induce GC formation. Mice were sacrificed after 10 days and spleens were isolated from control and SIRT3 knockout mice. The spleen sections derived from Sirt3^+/+ and Sirt3^−/−mice were stained by hematoxylin and eosin (FI&E) and peanut agglutinin (PNA) using standard procedures. The number of GCs, the total spleen area occupied by GCs and the average area occupied by the GCs were quantified using ImageJ 1,44o (NIH) software. To determine the percentage of GC B-cell population, single-cell suspensions from spleens derived from Sirt3^+/+ and Sirt3^−/− mice were stained using the following fluorescent-labeled anti-mouse antibodies: FITC conjugated anti-B220, PE conjugated anti-FAS, APC conjugated anti-CD38 from BD biosciences and analyzed by flow-cytometry. To evaluate follicular and marginal zone B-cell populations, splenic B cells were stained with APC conjugated anti-B220, FITC conjugated anti-CD21 and PE conjugated anti-CD38 from BD biosciences and then analyzed by flow-cytometry. DAPI was used for the exclusion of dead cells. The data was analyzed by FlowJo software.

Li M., Chiang Y.L., Lyssiotis C.A., Teater M.R., Hong J.Y., Shen H., Wang L., Hu J., Jing H., Chen Z., Jain N., Duy C., Mistry S.J., Cerchietti L., Cross J.R., Cantley L.C., Green M.R., Lin H, & Melnick A.M. (2019). Non-oncogene Addiction to SIRT3 Plays a Critical Role in Lymphomagenesis. Cancer cell, 35(6), 916-931.e9.

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Immunophenotyping of Cell Surface Receptors

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Cells were suspended in blocking buffer (0.5 % BSA/ 2 mM EDTA, in PBS) and then labeled with fluorochrome-conjugated antibodies: PE-conjugated anti-Fas (BD Biosciences, San Jose, CA, USA), PE-conjugated anti-FasL (Biolegend, San Diego, CA) (1 μg/ml), or anti-MHC-I (Biolegend, San Diego, CA), anti-RAE-1δ (Biolegend, San Diego, CA), anti-Rae-1αβγδε (scbt, CA, USA) (1:50), followed by APC-conjugated anti-mouse IgG (scbt, CA, USA) or APC-conjugated anti-rabbit IgG (scbt, CA, USA) (1:50) secondary antibodies, as appropriate. Cell membrane expression of these molecules was assessed by Flow Cytometry (FACS Aria III, BD Biosciences). Raw data was further analyzed by using Flow Jo software (Tree Star, Inc. Ashland OR).

Orozco-Morales M., Sánchez-García F.J., Golán-Cancela I., Hernández-Pedro N., Costoya J.A., de la Cruz V.P., Moreno-Jiménez S., Sotelo J, & Pineda B. (2015). RB mutation and RAS overexpression induce resistance to NK cell-mediated cytotoxicity in glioma cells. Cancer Cell International, 15, 57.

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Multiparametric Flow Cytometry Analysis

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FITC-conjugated anti-CD3, APC-conjugated anti-CD4, APC-conjugated anti-CD8, and their corresponding isotype controls were purchased from eBioscience (San Diego, CA). PE-Cy5-conjugated anti-CD4, PE-Cy5-conjugated anti-CD8, FITC-conjugated anti-CD44, and their corresponding isotype controls, were obtained from BioLegend (San Diego, CA). PE-conjugated anti-CD45RB, PE-conjugated anti-Fas, and their corresponding isotype controls were purchased from BD Bioscience (San Jose, CA). FITC-conjugated anti-CD28 and its corresponding isotype control were purchased from Serotec (Oxfordshire, UK). Rabbit polyclonal primary anti-Ki-67 and its isotype control were purchased from Abcam (Cambridge, UK) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively. PerCP-conjugated donkey anti-rabbit IgG antibody was obtained from Santa Cruz Biotechnology. T cell mitogen concanavalin A (Con A) was obtained from Sigma (St. Louis, MO).

Sheu T.T., Chiang B.L., Yen J.H, & Lin W.C. (2014). Premature CD4+ T Cell Aging and Its Contribution to Lymphopenia-Induced Proliferation of Memory Cells in Autoimmune-Prone Non-Obese Diabetic Mice. PLoS ONE, 9(2), e89379.

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T Cell Apoptosis Signaling Pathway

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FITC-conjugated anti-Fas, PE-conjugated anti-Fas, anti-FasL and isotype control (BD Biosciences, Mountain View, CA, USA) were used to analyze the expression of Fas and FasL on different T cell subsets according to the manufacturer's instructions. Activated caspase 8 was determined by flow cytometry using a commercially available activated caspase 8 detection kit (eBioscience) according to the manufacturer's instructions.
Enzyme-linked immunosorbent assay (ELISA) Concentrations of sFas and IL-10 in the serum were measured by enzyme-linked immunosorbent assay (ELISA) using commercial kits (R&D Systems, MN, USA).

Yang X., Sun B., Wang H., Yin C., Wang X, & Ji X. (2014). Increased serum IL-10 in lupus patients promotes apoptosis of T cell subsets via the caspase 8 pathway initiated by Fas signaling. Journal of Biomedical Research, 29(3), 232-240.

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