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Anti cd95

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Anti-CD95 is a monoclonal antibody that binds to the CD95 (Fas) receptor on the surface of cells. The CD95 receptor is involved in the regulation of programmed cell death or apoptosis. Anti-CD95 can be used in flow cytometry and other immunological applications to detect and quantify CD95-expressing cells.

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Lab products found in correlation

Anti cd3, by BD (4 mentions) Anti cd28, by BD (4 mentions) Anti cd8, by BD (3 mentions) Anti cd4, by BD (3 mentions) Anti cd138, by BD (3 mentions) Anti cd23, by Thermo Fisher Scientific (3 mentions) Anti cd138, by BioLegend (3 mentions) Anti cd4, by BioLegend (3 mentions) Anti gl7, by BD (2 mentions) Foxp3 fix perm buffer, by Thermo Fisher Scientific (2 mentions) Anti cd19, by BioLegend (2 mentions) Anti cd8, by Thermo Fisher Scientific (2 mentions) Anti cd4, by Thermo Fisher Scientific (2 mentions) Anti cd19, by BD (2 mentions) Anti cd11b, by BioLegend (2 mentions) Facscanto 2, by BD (2 mentions) Live dead fixable yellow dead cell stain, by Thermo Fisher Scientific (1 mentions) Anti ifn γ, by BD (1 mentions) Anti cd69, by BioLegend (1 mentions) Cd8 percp cy5, by BD (1 mentions)

Close spelling variants of this product

Anti-CD95-PE (8 citations) Anti-CD95-APC (6 citations) Anti-CD95 (Jo2) (4 citations) Anti-CD95 PE-Cy5 (4 citations) Anti-CD95-PE-Cy7 (4 citations) Anti-CD95-FITC (3 citations) Anti-CD95 [DX2] (3 citations) PE-conjugated anti-CD95 antibody (3 citations) PE anti-human CD95 mAb (2 citations) Anti-CD95 PE-Cy7 (clone Jo.2 (2 citations)

10 protocols using anti cd95

1

Comprehensive Flow Cytometry Analysis of Murine Splenocytes

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Flow cytometry analysis of spleen cell suspensions was performed using the following fluorescent-labeled anti–mouse antibodies: APC-conjugated anti-B220 (BD Biosciences #553092), anti-CD138 (BD Biosciences #558626) and anti-IgM (eBioscience #17–5790-82); PE-conjugated anti-IgD (BD Biosciences #558597), anti-CD23 (eBioscience #5010271) and anti-CD95 (BD Biosciences #554258); PECy7-conjugated anti-CD21 (BioLegend #123420), anti-CD95 (BD Biosciences #557653), anti-GL7 (BD Biosciences # 561530) and anti-CD86 (BD Biosciences #560582); PEVio770-conjugated anti-B220 (Miltenyi Biotec #130–102-308); FITC-conjugated anti-CXCR4 (BD Biosciences #551967) and anti-IgG1 (ebioscience #11–4011-85); Brilliant Violet 421-conjugated anti-CD138 (BioLegend #142507); PerCP-Cy5.5-conjugated anti-GL7 (BioLegend #144610), anti-CD138 (BioLegend #142510) and anti-IgM (BD Biosciences #550881), APC-Cy7-conjugated anti-CD19 (ebioscience #47–0193-82), AlexaFluor647-conjugated anti-BLIMP1 (BD Biosciences #563643). Sytox blue (ThermoFisher Scientific) or DAPI was used for the exclusion of dead cells. Data was acquired on a BD FACSCanto II (BD Biosciences) and analyzed using FlowJo v10.1 software (TreeStar).
Dominguez P.M., Ghamlouch H., Rosikiewicz W., Kumar P., Béguelin W., Fontán L., Rivas M.A., Pawlikowska P., Armand M., Mouly E., Torres-Martin M., Doane A.S., Calvo Fernandez M.T., Durant M., Della-Valle V., Teater M., Cimmino L., Droin N., Tadros S., Motanagh S., Shih A.H., Rubin M.A., Tam W., Aifantis I., Levine R.L., Elemento O., Inghirami G., Green M.R., Figueroa M.E., Bernard O.A., Aoufouchi S., Li S., Shaknovich R, & Melnick A.M. (2018). TET2 deficiency causes germinal center hyperplasia, impairs plasma cell differentiation and promotes B-cell lymphomagenesis. Cancer discovery, 8(12), 1632-1653.
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2

Multiparametric Phenotyping of T Cell Subsets

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To define the memory vs. naïve subsets, the following antibodies were used: SP34–2 (CD3; Alexa700, BD Biosciences), L200 (CD4; AmCyan, BD Bioscienes), SK-1 (CD8; PerCP-cy-5.5, BD Biosciences), MAB11 (TNFα; FITC, eBioscience), B27 (IFNγ; APC, BD Pharmingen), FN50 (CD69; PE, BD Biosciences), CD28.2 (CD28; PE-TexasRed, BD Biosciences), 150503 (CCR7; R&D Systems), and SA (PacBlue, BD Biosciences). CCR7 was biotinylated using the EZ-link Malemide-PEO Solid Phase Biotinylation kit from Fisher Scientific, as per manufacturer instructions. For cell recognition and T cell response assays, the following antibodies were used: anti-CD3 (clone: SP34–2, Pacific Blue; BD Biosciences), anti-CD8 (clone: SK1, TruRed; BD Biosciences), anti-CD4 (clone: L200, PE-Cy7, BV395; BD Biosciences), anti-CD28 (clone: CD28.2, PE; BD Biosciences), anti-CD95 (clone: DX2, FITC; BD Biosciences), anti-CD69 (clone: FN50, ECD, Biolegend), anti-CCR7 (clone: 150503, Pacific Blue, R&D Systems), anti-IFNγ (clone: B27, FITC; BD Biosciences), anti-TNFα (clone: MAb11, Alexa 700; BD Biosciences), and LIVE/DEAD Fixable Yellow Dead Cell Stain (Life Technologies) was used to assess cell viability.
Malouli D., Gilbride R.M., Wu H.L., Hwang J.M., Maier N., Hughes C.M., Newhouse D., Morrow D., Ventura A.B., Law L., Tisoncik-Go J., Whitmore L., Smith E., Golez I., Chang J., Reed J.S., Waytashek C., Weber W., Taher H., Uebelhoer L.S., Womack J.L., McArdle M.R., Gao J., Papen C.R., Lifson J.D., Burwitz B.J., Axthelm M.K., Smedley J., Früh K., Gale M J.r., Picker L.J., Hansen S.G, & Sacha J.B. (2022). Cytomegalovirus Vaccine-induced Unconventional T cell Priming and Control of SIV Replication is Conserved Between Primate Species. Cell host & microbe, 30(9), 1207-1218.e7.
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3

Comprehensive Lymphocyte Phenotyping

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Lymphocytes isolated from peripheral blood, LN, jejunum, and rectum were quantified for their activation (CD38+, CD69+, and HLADR+), proliferation (Ki67+), and naïve/memory phenotype (CD28+ and CD95+). Cells were first stained with live/dead stain (Thermo Fisher Scientific, Eugene, OR, USA). After washing, cells were further stained with cocktail of monoclonal antibodies (mAbs) including anti-CD3, anti-CD4, anti-CD8, anti-CD28, anti-CD95, anti-CD38, anti-CD69, and anti-HLADR antibodies (BD Biosciences, San Jose, CA, USA; Table S2) as reported earlier. To detect proliferating Ki67+ cells, intracellular staining protocol was performed using BD Fix/Perm solutions as described earlier [26 (link)]. Cells were fixed with 1x BD stabilizing and fixative buffer. At least 20,000 events were collected by gating on lymphocytes from each sample, and the data were analyzed using FlowJo software, version 10.7.0. (Becton, Dickinson & Company, Franklin Lakes, NJ, USA).
Pahar B., Gray W., Fahlberg M., Grasperge B., Hunter M., Das A., Mabee C., Aye P.P., Schiro F., Hensley K., Ratnayake A., Goff K., LaBranche C., Shen X., Tomaras G.D., DeMarco C.T., Montefiori D., Kissinger P., Marx P.A, & Traina-Dorge V. (2022). Recombinant Simian Varicella Virus-Simian Immunodeficiency Virus Vaccine Induces T and B Cell Functions and Provides Partial Protection against Repeated Mucosal SIV Challenges in Rhesus Macaques. Viruses, 14(12), 2819.
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4

Multiparameter Immunophenotyping Protocol

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The following antibodies were used for surface staining at 4°C for 30 min: anti-CD4 (Biolegend, RM4-5), anti-ICOS (Biolegend, 15F9), anti-CD19 (Biolegend, 6D5), anti-PD-1 (Biolegend, RMP1-30), anti-CXCR5 biotin (BD Biosciences, 2G8), T- and B-cell activation antigen (BD Biosciences, GL-7), CD38 (Biolegend, 90), CD138 (Biolegend, 281-2), and anti-CD95 (BD Biosciences, Jo2), Streptavidin-BV421 (Biolegend, 405225). For intracellular staining, samples were fixed with the Foxp3 Fix/Perm buffer set according to the manufacturers instructions (eBioscience). Samples were then intracellularly stained with anti-FoxP3 (eBiosciences, FJK-16S). No viability dye was included. Samples were analyzed on FACS Canto II (BD) or Cytek Aurora. Data was analyzed using FlowJo v10 (FlowJo LLC).
Cavazzoni C.B., Hanson B.L., Podestà M.A., Bechu E.D., Clement R.L., Zhang H., Daccache J., Reyes-Robles T., Hett E.C., Vora K.A., Fadeyi O.O., Oslund R.C., Hazuda D.J, & Sage P.T. (2022). Follicular T cells optimize the germinal center response to SARS-CoV-2 protein vaccination in mice. Cell Reports, 38(8), 110399.
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5

Detection of huCD4+ CAR Modified Cells

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To detect huCD4+ CAR modified cells, PBMCs and tissue necropsy samples were stained with the following antibodies: anti-human specific CD4 antibody (for detection and analysis of CD4CAR modified cells; Beckman Coulter, clone 13B8.2), anti-NHP CD45 (BD Biosciences, clone D058-1283), anti-CD4 (eBiosciences, clone OKT4), anti-CD8 (eBiosciences, clone SK1), anti-CD20 (eBiosciences, clone 2H7), anti-NK2Ga (Beckman Coulter, clone A60797), anti-CD14 (Beckman Coulter, clone IM2707U), anti-CD95 (BD Biosciences, clone DX2), anti-CD28 (BD Biosciences, clone D28.2), and anti-CD3 (BD Biosciences, clone SP34-2). Fluorophore conjugates included FITC, PE, PE-Cy5, PE-Cy7, alexa700, V500, efluor450, APC, and APC-efluor780.
Zhen A., Peterson C.W., Carrillo M.A., Reddy S.S., Youn C.S., Lam B.B., Chang N.Y., Martin H.A., Rick J.W., Kim J., Neel N.C., Rezek V.K., Kamata M., Chen I.S., Zack J.A., Kiem H.P, & Kitchen S.G. (2017). Long-term persistence and function of hematopoietic stem cell-derived chimeric antigen receptor T cells in a nonhuman primate model of HIV/AIDS. PLoS Pathogens, 13(12), e1006753.
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6

Multiparametric Flow Cytometry of B Cells

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For FC staining, non-specific binding was blocked using anti-FcR clone 2.4G2 and dead cells were excluded using cell viability dye (Tonbo Biosciences). The Abs were either purified in our lab or purchased and are as follows. Anti-B220 (Biolegend, RA3-6B2), anti-CD19 (BD ID3), anti-IgM (homemade, B7-6), anti-CD45 (home-made 30-F11), anti-CD21 (homemade, 7G6), anti-CD23 (ebioscience, B3B4) for B cells, anti-CD4 (Biolegend, GK1.5), anti-TCR-β (Biolegend H57-597) for T cells, anti-CD138 (Biolegend, 281-2) and anti-CD44 (Biolegend, IM7) for B cell blasts and PB, anti-CD73 (Biolegend, TY-11.8), anti-CD80 (ebioscience, 16-1oA1), anti-PD-L2 (ebioscience, TY25) for MBC, PNA (vector labs), anti-CD95 (BD, Jo2) for GCBC, anti-CD169 (Biolegend, 3D6.112) for metalophillic macrophages, anti-CD11b (Biolegend M1-70), anti-CD11c (ebioscience, N418), anti-CD69 (ebioscience, H1.2F3), anti-AID (ebioscience, mAID-2) and anti-T-bet (Biolegend, 4B10). The click IT Plus Edu kit was purchased from Invitrogen and the staining was done according to the recommended protocol. The cells were analyzed on LSR II or Fortessa instruments (BD) and the data were analyzed on FlowJo software.
Trivedi N., Weisel F., Smita S., Joachim S., Kader M., Radhakrishnan A., Clouser C., Rosenfeld A., Chikina M., Vigneault F., Hershberg U., Ismail N, & Shlomchik M. (2019). Liver is a generative site for the B cell response to Ehrlichia muris. Immunity, 51(6), 1088-1101.e5.
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7

Flow Cytometric Analysis of Spleen Immune Cells

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Spleens were harvested and single cell suspensions prepared using 100 µm filters. Flow cytometry for B and T cells was performed as previously described35 (link). Anti-CD19, anti-B220, anti-CD21, anti-CD23, anti-IgM, anti-CD3, anti-CD4, anti-CD8, anti-IL-17A, anti-IL-4 and anti-IFN-γ antibodies were obtained from eBioscience (San Diego, CA, USA), anti-IgD, anti- CD95, anti-CD138, anti-IgG1 and anti-IgG2ab antibodies from BD BioSciences, anti-IL-10 antibody was purchased from Biolegend (San Diego, CA, USA) and biotinylated peanut agglutinin (PNA) from Sigma-Aldrich (St Louis, USA).
Samples were measured on a FACS Canto II HTS or a LSR II flow cytometer (BD BioSciences) and analysis was performed using FlowJo v7.6 research software (Tree Star Inc. Ashland, OR).
Corneth O.B., Schaper F., Luk F., Asmawidjaja P.S., Mus A.M., Horst G., Heeringa P., Hendriks R.W., Westra J, & Lubberts E. (2019). Lack of IL-17 Receptor A signaling aggravates lymphoproliferation in C57BL/6 lpr mice. Scientific Reports, 9, 4032.
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8

Quantification of T Cell Subsets

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The numbers of circulating CD4+ T and CD8+ T lymphocytes were determined using BD Trucount absolute count tubes according to the manufacturer’s instructions (BD Biosciences). For detection of central memory CD4+ T (CD3 + CD4 + CD28 + CD95 + cells), effector memory CD4+ T (CD3 + CD4 + CD28-CD95+ cells) and TFH cell (CD3 + CD4 + CXCR5 + PD-1+) cells, PBMCs were stained with the following fluorescein-labeled antibodies: anti-CD3 (BD Bioscience), anti-CD4 (BD Bioscience), anti-CD8 (BD Bioscience), anti-CD28 (BD Bioscience), anti-CD95 (BD Bioscience), anti-CXCR5 (eBioscience), and anti-PD-1 (eBioscience) for 30 min and detected with a BD FACS LSR Fortessa flow cytometer (BD Biosciences, USA). Data were analyzed using FlowJo software (Tree Star, USA).
Li P., Wang Q., He Y., Yang C., Zhang Z., Liu Z., Liu B., Yin L., Cui Y., Hu P., Liu Y., Zheng P., Wang W., Qu L., Sun C., Guan S., Feng L, & Chen L. (2023). Booster vaccination is required to elicit and maintain COVID-19 vaccine-induced immunity in SIV-infected macaques. Emerging Microbes & Infections, 12(1), e2136538.
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9

Multiparametric Flow Cytometry Analysis

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The following antibodies were used for surface staining at 4°C for 30 minutes: anti-CD45 (Biolegend, 30-F11), anti-CD4 (Biolegend, RM4-5), anti-CD19 (Biolegend, 6D5), T- and B-cell activation antigen (BD Biosciences, GL-7), CD38 (Biolegend, 90), CD138 (Biolegend, 281-2), and anti-CD95 (BD Biosciences, Jo2). Samples were then intracellularly stained with anti-IgG1 (eBiosciences, FJK-16S) after fixation with the Foxp3 Fix/Perm buffer set according to the manufacturer’s instructions (eBioscience). Flow cytometry analysis was performed on a Cytek Aurora spectral analyzer and analyzed with FlowJo v10 (FlowJo LLC).
Zhang H., Cavazzoni C.B., Hanson B.L., Bechu E.D., Podestà M.A., Azzi J., Blazar B.R., Chong A.S., Kreisel D., Alessandrini A, & Sage P.T. (2022). Transcriptionally Distinct B Cells Infiltrate Allografts After Kidney Transplantation. Transplantation, 107(2), e47-e57.
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10

Multiparametric Flow Cytometry Analysis

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The following antibodies were used: Anti-CD3 (Biolegend 100237), anti-CD11b, (Biolegend 101237), anti-IgM (Biolegend 406525), anti-CD19 (BD Biosciences 561739), anti-CD95 (BD Biosciences 561856), anti-GL7 (BD Biosciences 553666), anti-CD21 APC (BD Biosciences 561770), anti-CD138 (BD Biosciences 553714), Horseradish peroxidase conjugated anti-mouse Light Chain Specific goat polyclonal antibody (Jackson Immunoresearch #115-035-174), anti-IgG1 (BD Biosciences 560089), and for histology anti-CD45R/B220 (Bio-Rad, MCA1258G) and anti-CD138 (Biolegend 142502).
McBride K.M., Kil H., Mu Y., Plummer J.B., Lee J., Zelazowski M.J., Sebastian M., Abba M.C, & Aldaz C.M. (2019). Wwox Deletion in Mouse B Cells Leads to Genomic Instability, Neoplastic Transformation, and Monoclonal Gammopathies. Frontiers in Oncology, 9, 517.
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