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Cd95 pe

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CD95-PE is a laboratory analysis product that detects the presence and levels of the CD95 protein, also known as Fas, on the surface of cells. It is a fluorescent-labeled antibody that binds specifically to the CD95 antigen. The core function of CD95-PE is to enable the identification and quantification of CD95-expressing cells through flow cytometry or other analytical techniques.

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Lab products found in correlation

Cd4 pe, by BD (2 mentions) Cd69 fitc, by BD (2 mentions) Cd8 pe, by BioLegend (2 mentions) Cd20 apc cy7, by BioLegend (2 mentions) Cd19 percp cy5, by BD (2 mentions) Cd19 fitc, by BioLegend (2 mentions) Cd38 fitc, by BioLegend (2 mentions) Cd25 bv605, by BioLegend (2 mentions) Cd80 pe cy7, by BioLegend (2 mentions) Mhcii apc, by BioLegend (2 mentions) Cd86 bv421, by BD (2 mentions) Cd40 pe dazzle, by BD (2 mentions) Cd14 pe cf594, by BioLegend (2 mentions) Cd27 pacificblue, by BioLegend (2 mentions) Zombie dye, by BioLegend (2 mentions) Gl7 fitc, by BD (2 mentions) Cd86 pe, by BioLegend (1 mentions) Cd80 percp cy5, by BioLegend (1 mentions) Mhcii pacificblue, by BioLegend (1 mentions) Cd40 pe, by BioLegend (1 mentions)

Close spelling variants of this product

PE-conjugated anti-CD95 antibody PE-Cy7-conjugated anti-CD95 CD95-PE-Cy5 (clone DX2), CD95/Fas-PE Anti-CD95 (Fas)-PE-Cy7 Anti-CD95-PE Anti-CD95-PE-Cy7 CD95‐PE‐CF594 Anti-human CD95-PE.Cy7 CD95-PE (DX2,

14 protocols using cd95 pe

1

Multiparametric Flow Cytometry of Murine Immune Cells

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Murine spleens and lymph nodes were dissected and passed through 70 µm strainers. After pre‐incubation with Zombie™ dye and Fc‐block (BioLegend), cells were stained with antibodies against B220‐FITC, B220‐PE‐Cy7, CD19‐FITC, CD25‐APC, CD69‐PerCP‐Cy5.5, CD95‐PE (all BD Bioscience), CD4‐PE, CD8‐FITC, CD11b‐BV510, CD21‐FITC, CD23‐APC, CD40‐PE, CD80‐PerCP‐Cy5.5, CD86‐PE and MHC‐II‐PacificBlue (all BioLegend). T cells were stained for interferon (IFN)‐γ‐APC (BioLegend) and IL‐17A‐PE (BD Bioscience) after 4‐h incubation with 50 ng/mL PMA, 0.5 μg/mL ionomycin and 1 μL/mL Golgi‐Plug; regulatory T cells were determined by intracellular staining for Foxp3‐PE (BD Bioscience).
Traub J., Traffehn S., Ochs J., Häusser‐Kinzel S., Stephan S., Scannevin R., Brück W., Metz I, & Weber M.S. (2019). Dimethyl fumarate impairs differentiated B cells and fosters central nervous system integrity in treatment of multiple sclerosis. Brain Pathology, 29(5), 640-657.
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2

T-cell Phenotyping by Flow Cytometry

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T cells were stained for cell-surface markers to differentiate T-cell lineage. Previous work (24 (link)) has demonstrated that CCR7, CD62L, CD45RO, and CD95 can be used to differentiate the various T-cell phenotypes by using the following expression patterns: naïve (TN)—CCR7+, CD62L+, CD45RO−, CD95−; stem central memory (TSCM)—CCR7+, CD62L+, CD45RO−, CD95+; central memory (TCM)—CCR7+, CD62L+, CD45RO+, CD95+; effector memory (TEM)—CCR7−, CD62L−, CD45RO+, CD95+; terminal effector (TEff)—CCR7−, CD62L−, CD45RO−, CD95+. The antibodies used for this analysis as well as quantitation of T-cell bulk described above were CD8-FITC (BD Biosciences; #347313), CD3-PE (BD Biosciences; #555340), CD4-APC (BD Biosciences; #555349), CCR7-FITC (BD Biosciences; #561271), CD95-PE (BD Biosciences; #556641), CD45RO (BD Biosciences; #559865), and CD62L-PE/Cy7 (BioLegend; 304822). Samples were then washed twice, and flow cytometry acquisition was performed on a BD FACSVerse Flow Cytometer or BD Accuri C6 (BD Biosciences).
Das R.K., Vernau L., Grupp S.A, & Barrett D.M. (2019). Naïve T-cell Deficits at Diagnosis and after Chemotherapy Impair Cell Therapy Potential in Pediatric Cancers. Cancer discovery, 9(4), 492-499.
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3

Multiparametric Flow Cytometry Analysis of Immune Cells

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Peripheral blood cells were stained with the following mAbs two times a week after transplantation: CD56 PE (B159), CD95 PE (DX2), HLA-I PE (DX17), CD3 PerCP (SP34–2), CD4 BV510 (L200), CD4 APC (L200) (BD Biosciences, San Jose, CA); CD8 APC (BW135.80), CD20 APC-Vio770 (LT20), CD25 Pacific Blue (BC96) (BioLegend), CD45RA APC-Vio770 (T6D11), CD11b Viogreen (M1/70.15.11.5), FOXP3 (236A/E7) (Invitrogen), HLA-I PE-Vio770 (REA230) (Miltenyi Biotec Inc. San Diego, CA); CD28 BV421 (CD28.2) and CD8 BV421 (RPA-T8), (BioLegend Inc. San Diego, CA). Anti-Bw6 FITC (H38, One Lambda, Canoga Park, CA) was used to assess donor chimerism. Tregs were defined as CD3+/CD4+/CD25high/FoxP3+. The same analysis was performed on liver tissue and lymph nodes on sacrifice. Data was acquired on a FACS Canto II Flow Cytometer (BD Bioscience) and analyzed using FloJo Software (TreeStar Inc. Ashland, OR).
Chaudhry S., Kato Y., Weiner J., Alonso-Guallart P., Baker S., Woodland DC I.V., Lefkowitch J.H., Duran-Struuck R., Sondermeijer H.P., Zitsman J., Sears M.L., Wu A., Karolewski B., Houck P.J., Martinez M., Kato T., Sykes M, & Griesemer A.D. (2020). Transient mixed chimerism with nonmyeloablative conditioning does not induce liver allograft tolerance in nonhuman primates. Transplantation, 104(8), 1580-1590.
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4

Multiparametric Flow Cytometry of Immune Cells

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After Zombie™ dye and Fc‐block (both BioLegend, San Diego, CA, USA) pre‐incubation, PBMC were stained with anti‐human CD14‐FITC, CD19‐PerCP‐Cy5.5, CD40‐PE‐Dazzle, CD69‐FITC, CD86‐BV421, CD95‐PE (all BD Bioscience, NJ, USA), CD4‐PE‐Cy7, CD8‐PE, CD14‐PE‐CF594, CD19‐FITC, CD20‐APC‐Cy7, CD24‐PerCp‐Cy5.5, CD25‐BV605, CD27‐PacificBlue, CD38‐FITC, CD80‐PE‐Cy7 and major histocompatibility complex (MHC)‐II‐APC (all BioLegend, CA, USA). Cytokines were evaluated after adding 1‐µL Golgi‐Plug (BioLegend) to CpG‐stimulated cells for 4 h; 500 ng/mL ionomycin and 20 ng/mL phorbol 12‐myristate 13‐acetate (PMA; Sigma Aldrich, MO) were added after 2 h. Cells were stained with anti‐human interleukin‐ (IL‐)6‐FITC, tumor necrosis factor (TNF)‐A700 and IL‐10‐PE‐CF594 (all BioLegend).
Traub J., Traffehn S., Ochs J., Häusser‐Kinzel S., Stephan S., Scannevin R., Brück W., Metz I, & Weber M.S. (2019). Dimethyl fumarate impairs differentiated B cells and fosters central nervous system integrity in treatment of multiple sclerosis. Brain Pathology, 29(5), 640-657.
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5

Apoptosis Analysis of Activated T Cells and DCs

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The cells were harvested on day 5 of MLR, and apoptosis was determined by staining cells with a combination of Annexin-V FITC with propidium iodide using the Annexin-V FITC Apoptosis Detection Kit (BD Pharmingen, USA), according to the manufacturers’ protocols. The cells were co-stained with CD3-APC. Additionally, cells were labelled with CD4-FITC (clone: RPA-T4, eBioscience, USA), CD8-APC (clone: RPA-T8, eBioscience, USA), CD95-PE (clone: DX2, BD Pharmingen, USA) or CD95L-PE (clone: NOK-1, BioLegend, USA) to identify T cells and CD11c-APC, CD95L-PE, Lineage Cocktail 1 (lin 1; CD3, CD14, CD16, CD19, CD20, CD56)-FITC (BD Pharmingen, USA) to identify DCs. The results were acquired and analyzed as described above.
Machcińska M., Kotur M., Jankowska A., Maruszewska-Cheruiyot M., Łaski A., Kotkowska Z., Bocian K, & Korczak-Kowalska G. (2021). Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms. Archivum Immunologiae et Therapiae Experimentalis, 69(1), 27.
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6

T-B Cell Activation and Phenotyping

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Day 3 primary T cells and day 1 primary B cells were incubated with peptide and anti-CD40 as described above in 1:1 and 1:10 T/B cell ratios at a total density of 5 × 106 cells per/mL. Three days later they were stained with anti CD4 allophycocyanin (ebioscience clone GK1.5), ICOS PE (ebioscience clone 7E.17G9) and PD1 FITC (ebioscience clone J43) for T-cell marker analysis. CXCR5 bio (BD clone 2G8) was used in combination with streptavidin PerCP (BD). For B-cell staining, GL7 FITC (BD), CD95 PE (BD) and CD19 allophycocyanin (BD, clone 1D3) were used.
Sinai P., Dozmorov I.M., Song R., Schwartzberg P.L., Wakeland E.K, & Wülfing C. (2014). T/B cell interactions are more transient in response to weak stimuli in SLE-prone mice. European journal of immunology, 44(12), 3522-3531.
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7

Comprehensive Profiling of T Cell Markers

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Cell surface proteins on (CAR-)T cells were stained with the following antibodies: annexin V-PE, CCR7-PerCP/Cy5.5, CD3-allophycocyanin (APC), CD3-BV421, CD4-APC/H7, CD8a-PE/Cy7, CD25-PE/Cy7, CD69-fluorescein isothiocyanate (FITC), CD95-PE, CD127-PerCP/Cy5.5, CXCR3-AF488 (all from BD Biosciences, San Jose, CA); CD8a-APC, CD8a-FITC, CD27-PE, CD28-FITC, CD45RA-APC, LAG-3-PE, TIGIT-PerCP/Cy5.5, TIM-3-FITC (all from BioLegend, San Diego, CA); and PD-1-APC (eBioscience, Thermo Fisher Scientific). Monocytes were stained with CD14-PE (eBioscience). To determine CAR expression, cells were labeled with a recombinant Fc-tagged BCMA protein (Enzo Life Sciences, Farmingdale, NY) followed by a BV421-conjugated anti-Fc antibody (BioLegend). Samples prepared from mouse organs were treated with Fc block (BD Biosciences) prior to staining. Cells were washed and resuspended in fluorescence-activated cell sorting (FACS) buffer (1% FCS in PBS) or 1% paraformaldehyde prior to acquisition. In experiments to assess viability and/or apoptosis, cells were resuspended in FACS buffer containing 1 μg/mL 7-aminoactinomycin D (7-AAD). In all other experiments, live cells were gated based on forward and side scatter.
All flow cytometry data were acquired on a FACSCanto II (BD Biosciences) and analyzed using FlowJo software version X.0.7 (Treestar, Ashland, OR).
Battram A.M., Oliver-Caldés A., Suárez-Lledó M., Lozano M., Bosch i Crespo M., Martínez-Cibrián N., Cid J., Moreno D.F., Rodríguez-Lobato L.G., Urbano-Ispizua A, & Fernández de Larrea C. (2022). T cells isolated from G-CSF-treated multiple myeloma patients are suitable for the generation of BCMA-directed CAR-T cells. Molecular Therapy. Methods & Clinical Development, 26, 207-223.
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8

Flow Cytometry Immunophenotyping Protocol

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The following antibodies were used for flow cytometry: CD3 Alexa 700 (SP34-2 BD Biosciences), CD4 AmCyan (L200 BD Biosciences), CD8 PerCP-Cy5.5 (SKI eBiosciences), CD8 AmCyan (SKI BD Biosciences), CD28 PE-Texas Red (CD28.2 Beckman Coulter, BD Biosciences), CD95 PE (DX2 BD Biosciences, eBiosciences), CCR5 Allophycocyanin (3A9 BD Biosciences), Ki-67 FITC (B56 BD Biosciences), CD56 PerCP-Cy5.5 (MEM-188 Invitrogen), CD16 Pacific Blue (3G8 BD Biosciences, Biolegend), CD20 Allophycocyanin-Cy7 (L27 BD Biosciences), HLA-DR PE-Texas Red (TU36 Invitrogen, Immu357 Beckman Coulter), NKG2A PE (Z199 Beckman Coulter), CD14 FITC (M5E2 BD Biosciences, R&D Systems), STAT5 PE (47/Stat5 (pY6) BD Biosciences), BrdU FITC (B44 BD Biosciences), BrdU Allophycocyanin (B44 BD Biosciences). Anti-CCR7 (150503) was purchased as purified immunoglobulin from R&D Systems, conjugated to biotin using a Pierce Chemical Co. biotinylation kit, and visualized with streptavidin–Pacific Blue (Invitrogen). Rhesus recombinant anti-IL-15 and rhesus recombinant control IgG1 mAb were provided through the National Institutes of Health’s Nonhuman Primate Reagent Resource Program. Simian recombinant IL-7 was provided by Cytheris SA (Issy-Les-Moulineaux, France). Rhesus recombinant IL-15 was provided by Francois Villinger (Emory University) through the Resource for Nonhuman Primate Immune Reagents.
DeGottardi M.Q., Okoye A.A., Vaidya M., Talla A., Konfe A.L., Reyes M.D., Clock J.A., Duell D.M., Legasse A.W., Sabnis A., Park B.S., Axthelm M.K., Estes J.D., Reinmann K.A., Sekaly R.P, & Picker L.J. (2016). Effect of anti-IL-15 administration on T cell and NK cell homeostasis in rhesus macaques. Journal of immunology (Baltimore, Md. : 1950), 197(4), 1183-1198.
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9

Comprehensive Immune Cell Profiling

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Bone marrow, peripheral blood, and splenic B cells from cCD79 and wild type mice were analyzed using the following antibodies: hCD79A-PE (R&D Systems, FAB69201P); hCD79B-PE (abcam, ab33295); mCD79B-AF488 ([HM79], made in house); hCD79A-AF647 ([Curly-14], made in house); B220-BV786 (BD, 563894); B220-APC (BD, 103212); B220-PE (BD, 561878); CD43 ACP-Cy7 (BD, 562866); BP1-PE (BD, 553735); CD24-APC (BioLegend, 101814); IgD-FITC (BioLegend, 405704); IgM-BUV396 (BD, 564025); CD19-FITC (BioLegend, 152404); CD93-APC (BioLegend, 136510); CD23-FITC (BD, 561772); CD1d-PE (BD, 553846); CD95-PE (BD, 554258); GL7-FITC (BD, 553666); fab anti-mIgG (H+L)-AF647 (Jackson ImmunoResearch, 115–606-072); phospho-Syk (Y352)-PE (BD, 557881); PTEN-PE (BD, 560002); mouse IgG1 isotype control-PE (BD, 559320).
Wemlinger S.M., Parker Harp C.R., Yu B., Hardy I.R., Seefeldt M., Matsuda J., Mingueneau M., Spilker K.A., Cameron T.O., Larrick J.W., Getahun A, & Cambier J.C. (2022). Preclinical Analysis of Candidate Anti-Human CD79 Therapeutic Antibodies Using a Humanized CD79 Mouse Model. The Journal of Immunology Author Choice, 208(7), 1566-1584.
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10

Multiparametric Flow Cytometric Analysis

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Fresh total cells from spleens were isolated in PBS1×-3% fetal bovine serum (FBS) and stained for 20 min at 4°C with the following monoclonal antibodies at predetermined optimal dilutions: CD121b-BV421, CD19-PeCF594, CD4-V500, CD8a-AF700, Bcl6-APC, CXCR5-Biotin, GL7-e450, CD95 PE, Foxp3-AF488, PD-1-PE (BD Biosciences) or PE-Texas Red (PETR), streptavidin-APC or streptavidin-APC-Cy7 (BD Biosciences), GITR PETR (Miltenyi). CXCR5 staining was performed using biotinylated anti-CXCR5 for 30 min at 20°C followed by APC-or APC-Cy7-labeled streptavidin at 4°C. Intracellular detection of Foxp3 was performed on fixed and permeabilized cells using appropriate buffer (eBioscience), following the manufacturer's recommendations. Stained cells were run on CytoFLEX S cytometer (Beckman -Coulter) and analyzed using FlowJo software (TreeStar Inc.). Dead cells were excluded by forward/side scatter gating.
Belbezier A., Engeroff P., Fourcade G., Vantomme H., Vaineau R., Gouritin B., Bellier B., Brocheriou I., Tchitchek N., Graff-Dubois S, & Klatzmann D. (2024). Interleukin-1 regulates follicular T cells during the germinal center reaction. Frontiers in immunology, 15.
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