Single‐cell suspensions from spleen were obtained from tissues and stained for 20 min with indicated antibodies. For intracellular staining, T cells were stimulated with eBioscience Cell Stimulation Cocktail for 6 h at 37 °C in the presence of GolgiStop. Cells were then stained with the surface marker for 15 min on ice and permeabilized using Cytofix/Cytoperm for 30 min on ice. Permeabilized cells were resuspended in Perm/Wash buffer and stained with cytokine antibody for 20 min. FACS analysis was performed with BD LSRII Flow Cytometer (BD) or Attune Flow Cytometers (ThermoFisher Scientific), and data were analyzed by BD FACSDiva, Attune NxT Software, or Flowjo software. For specific T cell analysis, splenocytes were harvested and stained with anti‐CD4, anti‐CD11a, and anti‐CD49d antibodies. Anti‐Foxp3, anti‐CD4, anti‐CD3, anti‐CD25, anti‐CD8, anti‐IFN‐γ, anti‐CD11b, anti‐Gr1, anti‐Ly6C, anti‐Ly6G, anti‐CD11a, and anti‐CD49d antibodies were purchased from BioLegend, eBioscience or Invitrogen.
Anti cd11a
Manufactured by BioLegend
Anti-CD11a is a lab equipment product that is used to detect and measure the expression of the CD11a molecule on the surface of cells. CD11a is a subunit of the lymphocyte function-associated antigen-1 (LFA-1) integrin, which plays a crucial role in cell-cell adhesion and immune cell trafficking.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd11b, by BioLegend (3 mentions)
Anti ly6g, by BioLegend (3 mentions)
Anti cd3, by BioLegend (2 mentions)
Anti gr1, by BioLegend (2 mentions)
Anti cd8, by BioLegend (2 mentions)
Anti ifn γ, by BioLegend (2 mentions)
Facscanto 2, by BD (2 mentions)
Lsr 2 flow cytometer, by BD (1 mentions)
Cell stimulation cocktail, by Thermo Fisher Scientific (1 mentions)
Attune flow cytometer, by Thermo Fisher Scientific (1 mentions)
Anti foxp3, by BioLegend (1 mentions)
Anti cd49d, by BioLegend (1 mentions)
Anti cd4, by BioLegend (1 mentions)
Anti ly6c, by BioLegend (1 mentions)
Cellquest pro software, by BD (1 mentions)
Anti cd162 psgl 1, by BD (1 mentions)
Annexin 5 pi kit, by Thermo Fisher Scientific (1 mentions)
Anti stat1, by Cell Signaling Technology (1 mentions)
Anti p stat1 tyr701, by Cell Signaling Technology (1 mentions)
Anti icam 1, by BioLegend (1 mentions)
6 protocols using anti cd11a
1
Multiparameter Flow Cytometry Analysis
2
Kinetics of Immune Cell Response in K/BxN Serum Transfer Arthritis
3
Comprehensive Neutrophil Phenotyping and Signaling
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Harvested cells were stained with antibodies including: anti-Ly6G (1:200 dilution; BioLegend, no. 127606, 127610, or 127618), anti-CD11b (1:200 dilution; BioLegend, no. 101206), anti-CD11a (1:200 dilution; BioLegend, no. 153103), anti-SIRPα (1:200 dilution; BioLegend, no. 37210), anti-CD62L (1:200 dilution; BioLegend, no. 104428), anti-PD-L1 (1:200 dilution; BioLegend, no. 124312), anti-CXCR2 (1:300 dilution; BioLegend, no. 149604), anti-ICAM1 (1:300 dilution; BioLegend, no. 116121), anti-CD29 antibodies (1:300 dilution; BioLegend, no. 102221). In some analysis mentioned in the results, cells were treated with a fixation kit (BD Biosciences), subsequently stained with anti-STAT1 (1:100 dilution; Cell Signaling, no. 80916S), anti-p-STAT1 (Tyr701) (1:100 dilution; Cell Signaling, no. 8009S), anti-p-Src (Tyr416) (1:20 dilution; ThermoFisher, no. MA5-28055), or anti-Fyn (1:50 dilution; Santa Cruz, no. sc-434 FITC). Surface phenotype, transcription factor and intracellular protein levels of Ly6G+ neutrophils were analyzed using FACSCanto II (BD Biosciences). Neutrophil viability was assessed by staining and flow analysis with the annexin V/PI kit (1:4,000 dilution; Thermo Fisher Scientific, no. P3566) as described previously74 (link).
4
NK Cell Receptor Expression Analysis
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The following FITC‐, PE‐, PE‐Cy5–, or APC‐conjugated mAbs were obtained from BD Bioscience, eBioscience, or Biolegend: anti‐NKG2D, anti‐LFA‐1 (CD11a), anti–DNAM‐1 (CD226), anti‐MICA/B, anti‐ULBP1, anti‐ULBP2, anti‐ULBP4, anti‐ICAM‐1 (CD54), anti‐ICAM‐2 (CD102), anti‐PVR (CD155), and anti‐Nectin‐2 (CD112). Additionally, the following blocking mAbs were also used: anti‐NKG2D (1D11 from eBioscience), anti‐CD11a (HI111 from Biolegend), anti‐CD226 (DX11 from Abcam), and mouseIgG1κ isotype control (P3.6.2.8.1 from eBioscienc). Specific extracellular signal–regulatory kinase1/2 (ERK1/2) inhibitor (PD098059) was purchased from Sigma‐Aldrich.
5
Phenotypic Analysis of Virus-Specific CD8+ T Cells
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Mononuclear cells from blood, spleen and tonsils were stained with fluorochrome-conjugated antibodies (mAbs) specific for cell surface proteins. The following mAbs were used for identification of CD8+ T cells and the determination of their phenotype; anti-CD3, anti-CD8 (Biolegend), anti-CCR7 (R&D Systems), anti-CD45RA (BD Biosciences), anti-CD69, anti-CD25, anti-CD137, anti-HLA-DR, anti-CD103, anti-KLRG-1 and anti-CD11a (all obtained from Biolegend). Fluorochrome-conjugated HLA class I dextramers (Immudex) were used to identify virus-specific CD8+ T cells. EBV-specific CD8+ T cells were identified with dextramers specific for the following viral epitopes; GLCTLVAML (derived from EBV-lytic protein BMLF1), CLGGLLTMV (derived from EBV-latent protein LMP2), RAKFKQLL (derived from EBV-lytic protein BZLF1) and FLRGRAYGL (derived from EBV-latent protein EBNA3A). CMV-specific CD8+ T cells were identified with dextramers specific for the following viral epitopes; NLVPMVATV, TPRVTGGGAM, RPHERNGFTV (all derived from pp65 protein), VLEETSVML, ELRRKMMYM, ELKRKMMYM (all derived from IE-1 protein) and VTEHDTLLY (derived from pp50 protein). Stained cells were analyzed on either FACSCanto II or LSRFortessa flow cytometer (BD Biosciences) and the data processed using FlowJo software (Treestar, Ashland, USA).
6
Multiparametric analysis of tumor-infiltrating immune cells
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Single-cell suspensions were prepared from the tumor tissues. For cell-surface staining, all samples were incubated with a Zombie Aqua Fixable Viability Kit (BioLegend, cat. #423101) for 15 min at room temperature. To detect cytokine production, cells were treated with a Cell Activation Cocktail (BioLegend, cat. #423304) for 4 h. The cells were stained with anti-CD45 antibody (BioLegend, cat. #103108), anti-CD8a (BioLegend, cat. #100714), anti-CD45.2 (BioLegend; cat. #109829), anti-TCR Vβ5.1,5.2 (BioLegend, cat. #139508), anti-CD11a (BioLegend; cat. #101119), anti-CD11a/CD18 (BioLegend; cat. #141012), and anti-CD69 (BioLegend; cat. #104539). After fixation and permeabilization, cells were stained with anti-IFN-γ (BioLegend, cat. #505808) and anti-TNF-α (BioLegend; cat. #506322). For intranuclear detection, the cells were fixed and permeabilized using a Transcription Factor Buffer Set (BD eBioscience, cat. #51–9008102) and stained with anti-Ki67 (BioLegend, cat. #652410), or anti-STAT5 phospho (Tyr694) (BioLegend, cat. #936906). Data were acquired on a Beckman Coulter CytoFLEX flow cytometer and analyzed using FlowJo software.
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