Cd95 bv421

Manufactured by BioLegend

CD95-BV421 is a fluorescently labeled antibody that binds to the CD95 (Fas) protein. CD95 is a cell surface receptor that plays a role in the regulation of programmed cell death (apoptosis).

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Lab products found in correlation

Ifn α, by Merck Group (1 mentions) 7 aad, by BD (1 mentions) Cd19 pe lt19, by Miltenyi Biotec (1 mentions) Cd138 allophycocyanin b b4, by Miltenyi Biotec (1 mentions) Cd38 af700, by BioLegend (1 mentions) Cd20 efluor v450 2h7, by Thermo Fisher Scientific (1 mentions) Cd27 af647 lt27, by Bio-Rad (1 mentions) Cd27 fitc m t271, by BD (1 mentions) Cd19 percp cy5.5 sj225c1, by BD (1 mentions) Cd24 fitc ml5, by BD (1 mentions) Cd27 bv605, by BioLegend (1 mentions) Cd3viogreen bw264 56, by Miltenyi Biotec (1 mentions) Unconjugatedgoat anti irf4 m 17, by Santa Cruz Biotechnology (1 mentions) Donkey anti goat igg af488, by Thermo Fisher Scientific (1 mentions) Goat anti human f ab 2 fragments anti igm and igg, by Jackson ImmunoResearch (1 mentions) Hybridomax hybridoma growth supplement, by Gentaur (1 mentions) Mem amino acids solution, by Merck Group (1 mentions) Ki 67 fitc b56, by BD (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Il 21, by Thermo Fisher Scientific (1 mentions)

4 protocols using cd95 bv421

Multiparametric Flow Cytometry of B Cells

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Antibodies used were CD19 PE (LT19; Miltenyi), CD138 allophycocyanin
(B-B4; Miltenyi Biotec), CD38 PE-Cy7 (HB7; BD Biosciences), CD38 AF700 (HIT2;
Biolegend), CD20 efluor V450 (2H7; eBioscience); CD27 AF647 (LT27; AbD Serotec),
CD27 FITC (M-T271), CD19 PerCP-Cy5.5 (SJ225C1; BD Biosciences), CD19 PE-Cy7
(SJ225C1; BD Biosciences), CD24 FITC(ML5; BD Biosciences), CD84 PE (CD84.1.21;
Biolegend), CD38 PerCP-Cy5.5 (HIT2; BD Biosciences), CD95 BV421 (DX2;
Biolegend), CD20 APC-H7(L27; BD Biosciences), CD27 BV605 (O323; Biolegend), CD3
VioGreen (BW264/56; Miltenyi), Ki67 FITC (B56; BD Biosciences), unconjugated
goat anti-IRF4 (M-17; Santa Cruz) and donkey anti-goat IgG AF488 (Polyclonal;
Invitrogen). Controls were isotype-matched mouse mAbs. Annexin V FITC was from
eBioscience and 7-AAD from BD Biosciences.
Reagents included human IL-2 (Roche); IL-6 (Peprotech) and IFN-α
(Sigma); IL-21 (PeproTech); goat anti-human F(ab′)₂ fragments
(anti-IgM and -IgG; Jackson Immunoresearch); HybridoMax hybridoma growth
supplement (Gentaur); Lipid Mixture 1, chemically defined (200X) and MEM Amino
Acids Solution (50X; Sigma).

Arumugakani G., Stephenson S.J., Newton D.J., Rawstron A., Emery P., Doody G.M., McGonagle D, & Tooze R.M. (2017). Early Emergence of CD19-Negative Human Antibody-Secreting Cells at the Plasmablast to Plasma Cell Transition. The Journal of Immunology Author Choice, 198(12), 4618-4628.

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Multicolor Flow Cytometry of PBMC

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We isolated peripheral blood mononuclear cells (PBMC) from peripheral blood by Ficoll gradient and we stored them in liquid nitrogen for batched analysis. For multicolor flow cytometry analyses, we used the following monoclonal antibodies: from Becton Dickinson (BD, Franklin Lakes, NJ), CD3-FITC, CD3-PerCP-Cy5.5, CD4-APC-Cy7, CD8-BV510, CD45RO-FITC, CD45RA-APC, CD45RA-APC-Cy7, CD71-PE, IgD-PerCP-Cy5.5, CD25-APC-Cy7, CD138-BV421, CCR4-PE, CD27-PE, CD95-BV421, CD28-BV421, TIM3-BV421, CCR6-BV421, CCR7-AF700, CXCR3-PE, IL-2-PE, IL-17-BV786, and IFN-γ-PE-Cy7; from Biolegend (San Diego, CA), CD19-BV510, CD56-FITC, CD27-APC, CD127-FITC, CD57-PerCp-Cy5.5, PD-1-APC-Cy7, CXCR5-FITC, LAG3-APC, IL-10-PerCP-Cy5.5, and TNF-α-FITC. From eBioscience (San Diego, CA), CD4-PE-Cy7, CD21-PE, and CD24-APC-Cy7 were used. From Miltenyi Biotec (Auburn, CA), CD25-APC and anti-KLRG1-PE were used. From Beckman Coulter (Indianapolis, IN), CD38-PE-Cy7 was used.
We performed intracellular staining for IL-2, IL-4, IL-17, IFN-g, and TNF-a together with extracellular markers for CD4⁺, CD8⁺, and CD19⁺. Cells were fixed and permeabilized using Intracellular Fixation and Permeabilization Buffer Set (eBioscience) according to the manufacturer’s instructions.
Data were acquired (> 1 × 10⁶ events) on a 3-laser FACSLyric flow cytometer (BD Biosciences) and analyzed with FlowJo^® software.

Tawhari I., Hallak P., Bin S., Yamani F., Safar-Boueri M., Irshad A., Leventhal J., Ansari M.J., Cravedi P, & Gallon L. (2022). Early calcineurin-inhibitor to belatacept conversion in steroid-free kidney transplant recipients. Frontiers in Immunology, 13, 1096881.

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Multiparameter T Cell Phenotyping

Cited 1 time

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Blood samples were drawn from patients following permission and processed at the University of Arizona Biorepository Laboratory. Peripheral blood mononuclear cells were cryopreserved for future analysis. A complete blood count was performed using an A^c-T 5diff CP machine (Beckman Coulter, Pasadena, CA). Cryopreserved PBMC (1–2x10⁶/sample) were stained with LIVE/DEAD Fixable Dead Cell Stain-AQUA (Invitrogen) and T cell markers in various combinations.^{11 (link)} The following mAbs were used to differentiate T cell subsets: CD3—BV570 (BioLegend), CD4—APC (eBioscience), CD8β—ECD (Beckman Coulter), CD95—BV421 (BioLegend), CD28—PerCp/Cy5.5 (BioLegend), CCR7—FITC (BD Pharmogen), CD45RA—BV605 (BioLegend), CD57—BV570 (BioLegend), IFN-γ—APCe780 (eBioscience). Cells following various combination and incubations as described in our previous study,^{11 (link)} were analyzed on the BD LSR II instrument using DiVa acquisition (BDIS, Mountain View, CA) and the FlowJo analysis software (TreeStar Inc., Ashland, OR).
Twenty (20) of 37 patients in this study completed the entire cell surface analysis.

Klotz S.A., Egurrola C., Love M., Miller M.N., Bradley N., Smith S.N., Najafi B, & Ahmad N. (2022). Phenotypic frailty in people living with HIV is not correlated with age or immunosenescence. International Journal of STD & AIDS, 33(6), 597-603.

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Phenotyping and Transduction Efficiency

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To phenotype expanded cells, cells were stained with the following primary antibodies (from Miltenyi Biotec unless otherwise stated) CD3-PE Vio770, CD4-Vio Blue, CD8-PE, CD45-VioGreen, CD45RA -PE Vio770, CCR7-PE, CD62L-APCCy7 (Biolegend), CD95-BV421 (Biolegend), CD14-APC, CD20-APC Vio770, CD56-PE Vio770. To assess the efficiency of CD19-CAR transduction, cells were stained using a Biotin-SP (long spacer) AffiniPure F(ab') Fragment Goat Anti-Mouse IgG, F(ab') Fragment Specific antibody (Jackson Immunoresearch) followed by Streptavidin-APC or Streptavidin-FITC (Biolegend).
Cells were acquired on a 4-laser BD LSRII and FACS analysis performed using FlowJo v10.

Mock U., Nickolay L., Philip B., Cheung G.W., Zhan H., Johnston I.C.D., Kaiser A.D., Peggs K., Pule M., Thrasher A.J, & Qasim W. (2016). Automated manufacturing of chimeric antigen receptor T cells for adoptive immunotherapy using CliniMACS prodigy. Cytotherapy, 18(8).

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