Anti cd95 pe

Manufactured by BD

Sourced in United States

Anti-CD95-PE is a fluorescently labeled monoclonal antibody that binds to the CD95 (Fas) receptor on the surface of cells. It is used for the identification and quantification of CD95-expressing cells in flow cytometry applications.

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Lab products found in correlation

Streptavidin apc, by BD (4 mentions) Pe anti cd138, by BD (3 mentions) Anti b220 percp, by BD (3 mentions) Facscalibur, by BD (3 mentions) Pna biotin, by Vector Laboratories (2 mentions) Apc annexin 5 apoptosis detection kit with 7 aad, by BioLegend (2 mentions) Anti cd21 apc, by BioLegend (2 mentions) Igm apc clone 2 41, by BD (2 mentions) Anti igd pe clone 11 26c 2a, by BD (2 mentions) Anti cd23 pe clone b3b4, by BD (2 mentions) Anti cd24 hsa pe, by BioLegend (2 mentions) Anti cd93 pe, by BioLegend (2 mentions) Anti cd23 pacific blue, by BioLegend (2 mentions) Anti cd19 cf594, by BD (2 mentions) Anti cd3 bv605, by BioLegend (2 mentions) Anti pd 1 fitc, by BD (2 mentions) Anti ccr7 af700, by BD (2 mentions) Anti cd19 pacific blue, by Thermo Fisher Scientific (2 mentions) Anti cd27 pc5, by Beckman Coulter (2 mentions) Anti cd45ra pe cy7, by BD (2 mentions)

Close spelling variants of this product

CD95-PE (14 citations) Anti-CD95 (10 citations) PE-Cy7-conjugated anti-CD95 (4 citations) Anti-CD95-PE-Cy7 (4 citations) Anti-CD95 PE-Cy5 (4 citations) PE-conjugated anti-CD95 antibody (3 citations) Anti-CD95 (Fas)-PE-Cy7 (2 citations) Anti-human CD95-PE.Cy7 (2 citations) PE anti-human CD95 mAb (2 citations) Anti-CD95 PE-Cy7 (clone Jo.2 (2 citations)

8 protocols using anti cd95 pe

Flow Cytometric Immunophenotyping of Murine Immune Cells

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Bone marrow cells, splenocytes, and peritoneal wash cells were depleted of red blood cells and stained with various combinations of the following antibodies. Bone marrow: anti-CD43 FITC (BD Biosciences), anti-IgM PE (BD Biosciences), anti-B220 PerCP-Cy5.5 (Tonbo Biosciences), and anti-CD93 APC (Invitrogen). Spleen: anti-CD21 FITC (BD Biosciences), anti-CD23 PE (BD Biosciences), anti-CD95 PE (BD Biosciences), anti-IgM PErCP-Cy5.5 (BD Biosciences), B220 PerCP-Cy5.5, anti-B220 APC (Tonbo Biosciences), or anti-GL7 APC (BD Biosciences). Peritoneal wash: anti-CD11b FITC (BD Biosciences), anti-CD5 PE (Tonbo Biosciences), anti-IgM PErCP-Cy5.5, anti-B220 APC. Cultured B cells were stained with anti-CD138 PE (BD Biosciences), anti-B220 PerCP-Cy5.5, and anti-IgG1 biotin (BD Biosciences) plus streptavidin APC (Tonbo Biosciences). Samples were run on a FACS Calibur (BD) and analyzed with Flowjo (Treestar).

Ottens K., Schneider J., Kane L.P, & Satterthwaite A.B. (2020). PIK3IP1 promotes extrafollicular class switching in T-dependent immune responses. Journal of immunology (Baltimore, Md. : 1950), 205(8), 2100-2108.

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Splenic Immune Profiling in Infected Mice

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The spleens of mice were collected at various time points post-infection, as detailed in figure legends. Splenic cells were passed through a 70 mm cell strainer (Fisher Scientific), centrifuged, and then treated with Ammonium-Chloride-Potassium (ACK) red blood cell lysing buffer (Gibco). Following washes, 4 x 10⁶ total spleen cells were incubated with anti-mouse CD16/CD32 (Fc Block, BD Biosciences; Mississauga, ON, Canada) for 15 min on ice according to manufacturer’s instructions. Afterwards, surface staining with the following antibodies: anti-CD95^PE, anti-CD86^biotin, anti-CD19^PE-Cy7, anti-GL-7^AF647 and anti-CD184^BV421 (all BD Biosciences) was performed for 30 min on ice. Following washing, cells were incubated with Brilliant Stain Buffer (BD Biosciences) and stained with Streptavidine^BV605 (BD Biosciences) for 10 min on ice. Cells were then washed and immediately analyzed on a BD FACSaria Fusion. The data collected was analyzed on FlowJo version 10. All antibodies used are listed in S1 Table.

Dolbec D., Lehoux M., de Beauville A.A., Zahn A., Di Noia J.M, & Segura M. (2024). Unmutated but T cell dependent IgM antibodies targeting Streptococcus suis play an essential role in bacterial clearance. PLOS Pathogens, 20(1), e1011957.

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Quantifying GC B cells and Tfh cells

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To quantify germinal center (GC) B cells and follicular helper T-cells (Tfh) we stained isolated draining lymph node cells with anti-CD4 pacific blue, anti-B220 PercP, anti-PD-1 PE, anti-CXCR5 biotin, anti-CD95 PE, anti-GL7 FITC and streptavidin APC (all from BD Biosciences). One million events in a live lymphocyte gate were acquired on a FACSCanto flow cytometer (BD Biosciences) and then analyzed using FlowJo software (version 10, Tree Star). Germinal center B cells were identified as CD4^-B220⁺GL7⁺CD95⁺ and Tfh as CD4⁺B220^-PD1⁺CXCR5⁺ population. S1 Fig shows the gating strategy used to identify each cell population.

Apostólico J.D., Boscardin S.B., Yamamoto M.M., de Oliveira-Filho J.N., Kalil J., Cunha-Neto E, & Rosa D.S. (2016). HIV Envelope Trimer Specific Immune Response Is Influenced by Different Adjuvant Formulations and Heterologous Prime-Boost. PLoS ONE, 11(1), e0145637.

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Multiparametric Flow Cytometry Analysis of B Cell Subsets

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Spleen cell suspensions or cultured B-cells were stained with the following antibodies as described:^{25 (link),46 (link)} anti-CD138-PE (clone: 281-2); anti-CD95-PE (clone: Jo2); IgM-APC (clone II/41); anti-IgD-PE (clone 11-26c.2a); and anti-CD23-PE (clone B3B4) (all BD Pharmingen); and anti-B220-PerCP (clone: RA3-6B2); anti-CD21-APC (clone: 7E9); anti-CD24 (HSA)-PE (clone: 30-F1), anti-CD93-PE (clone: AA4.1); and anti-CD23-Pacific Blue (clone: B3B4) (all Biolegend); and anti-CD19-CF594 (clone: 1D3) (BD Horizon); and PNA-Biotin (Vector Laboratories) followed by Streptavidin-APC (BD Pharmingen). Annexin V/7-AAD stainings were performed using the APC Annexin V Apoptosis Detection Kit with 7-AAD (Biolegend). For DNA content analysis, cells were lysed and stained with propidium iodide (PI). The cells were analyzed on a FACSCalibur or a LSRII (Becton Dickinson). Transitional B-cells were identified by gating on B220⁺CD93⁺ lymphocytes.^{39 (link)} GC B-cells were identified by gating on B220⁺ lymphocytes. eGFP⁺ and eGFP⁻ CD138^hi plasma cells were identified through the lymphocyte gate. Data were analyzed using FlowJo software.

Milanovic M., Heise N., De Silva N.S., Anderson M.M., Silva K., Carette A., Orelli F., Bhagat G, & Klein U. (2016). Differential requirements for the canonical NF-κB transcription factors c-REL and RELA during the generation and activation of mature B-cells. Immunology and cell biology, 95(3), 261-271.

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Multiparametric Flow Cytometry Analysis of B Cell Subsets

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Multiparametric Flow Cytometry for Immune Profiling

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The following fluorochrome-conjugated monoclonal antibodies (mAbs) were purchased from commercial vendors and used for surface staining: anti-CD4-V500, anti-CD8-APC-H7, anti-CD45RA-PE-Cy7, anti-CD45RO-APC, anti-CCR7-AF700, anti-CD95-PE, anti-CD160-FITC, anti-CD244-FITC, and anti-PD-1-FITC (BD Biosciences, San Jose, CA); anti-CD3-BV605 (Biolegend, San Diego, CA); anti-CD14-Pacific Blue and anti-CD19-Pacific Blue (Life Technologies, Carlsbad, CA); and anti-CD27-PC5 (Beckman Coulter, Indianapolis, IN). The fixable violet amine reactive dye (ViViD; Invitrogen/Molecular Probes, Eugene, OR) was used to eliminate dead cells by flow cytometry. For intracellular cytokine staining, the following mAbs were used: anti-granzyme B (GZMB)-FITC, anti-IL-2-FITC, and anti-IFN-γ-FITC (BD Biosciences).

Hosokawa K., Muranski P., Feng X., Townsley D.M., Liu B., Knickelbein J., Keyvanfar K., Dumitriu B., Ito S., Kajigaya S., Taylor JG V.I., Kaplan M.J., Nussenblatt R.B., Barrett A.J., O’Shea J, & Young N.S. (2016). Memory Stem T Cells in Autoimmune Disease: High Frequency of Circulating CD8+ Memory Stem Cells in Acquired Aplastic Anemia. Journal of immunology (Baltimore, Md. : 1950), 196(4), 1568-1578.

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Multiparametric Flow Cytometry Analysis

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The following fluorochrome-conjugated monoclonal antibodies (mAbs) were purchased from commercial vendors and used for surface staining: anti-CD4-V500, anti-CD8-APC-H7, anti-CD45RA-PE-Cy7, anti-CD45RO-APC, anti-CCR7-AF700, anti-CD95-PE, anti-PD-1-FITC and anti-CD69-FITC (BD Biosciences, San Jose, CA); anti-CD3-BV605 (Biolegend, San Diego, CA); anti-CD14-Pacific Blue and anti-CD19-Pacific Blue (Life Technologies, Carlsbad, CA); anti-CD30 Ligand /TNFSF8-AF488 and anti- A20/TNFAIP3- AF488 (Novus Biologicals, Littleton, CO) and anti-CD27-PC5 (Beckman Coulter, Indianapolis, IN). The fixable violet amine reactive dye (ViViD; Invitrogen/Molecular Probes, Eugene, OR) was used to eliminate dead cells by flow cytometry.

Hosokawa K., Kajigaya S., Keyvanfar K., Qiao W., Xie Y., Townsley D.M., Feng X, & Young N.S. (2017). T Cell Transcriptomes from Paroxysmal Nocturnal Hemoglobinuria Patients Reveal Novel Signaling Pathways. Journal of immunology (Baltimore, Md. : 1950), 199(2), 477-488.

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Multicolor Flow Cytometry Analysis

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CRC lines, cells separated from human CRC tissue fragments, DCs and effector leukocytes (mentioned in previous paragraphs) were stained with the following cocktail of monoclonal antibodies purchased from BD Biosciences, USA: anti-CD29-APC (clone MAR4, IgG1κ), anti-CD44-FITC (clone C26, IgG2bκ), anti-CD95-PE (clone DX2, C3H/Bi IgG1κ), anti-FasL Biotin (clone NOK-1, IgG1) coupled with Streptavidin-APC, anti-CD3-PE (clone UCHT1, IgG1κ), anti-CD4-FITC (clone PA-T4, IgG1κ), anti-CD11c-APC (clone S-HCL-3, IgG2b),anti-CD14-PerCP (clone MøP9, IgG2b), anti-CD25-PE (clone M-A251, IgG1κ), anti-CD56 (clone B159, IgG1κ), anti-CD80-PE (clone L307, IgG1κ), anti-CD83-APC (clone HB15e, IgG1κ), anti-HLA-DR-PerCP (clone L243, IgG2a). Anti-CD133/2-PE (clone 293C3, IgG2bκ) monoclonal antibodies were purchased from MiltenyiBiotec. After 30 min incubation in the dark, samples were fixed with 1% PFA or PBS + 1 mM EDTA for adherent or spherical cells, respectively, and prepared for further analysis. Flow cytometric analysis was performed using FACS Calibur flow cytometer (BD Biosciences, USA) with BD CellQuest Pro Software.

Szaryńska M., Olejniczak A., Kobiela J., Łaski D., Śledziński Z, & Kmieć Z. (2018). Cancer stem cells as targets for DC-based immunotherapy of colorectal cancer. Scientific Reports, 8, 12042.

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