Cx3cr1gfp/+ mice were infected with 2x108 Yptb-OVA, and on day 6 post infection LP cells were isolated from the ileum and cecum. Cells were stained with antibodies for CD11c, CD11b, I-Ab and dump gate (CD103, IgA (AD3, LSBio), CD19 (1D3, eBioscience), TCRβ (H57-597, eBioscience), TCRγδ (GL3, eBioscience)) sorted using a FACSAria (BD) to greater than 96% purity. Congenically marked CD8+CD44hi memory cells were sorted from Yptb-OVA memory mice and labelled with CFSE. 5x104 APCs and 5x104 memory T cells were mixed and incubated for 3 days with the addition of 5U/ml IL-2 for the final day of incubation. CFSE dilution was examined by flow cytometry and a live/dead stain was used to exclude dead cells from the analysis. Yptb- OVA memory cells were also incubated with APCs pulsed with SIINFEKL and YopE69-77 peptides to determine the maximum antigen specific response, and 60-70% of sorted Yptb-OVA memory cells divided in response to this peptide mixture.
Tcrγδ gl3
Manufactured by Thermo Fisher Scientific
The TCRγδ (GL3) is a monoclonal antibody that binds to the gamma-delta T cell receptor. It is commonly used for the identification and analysis of gamma-delta T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Tcrβ h57 597, by Thermo Fisher Scientific (4 mentions)
Cd19 1d3, by Thermo Fisher Scientific (3 mentions)
Facsaria, by BD (2 mentions)
Iga ad3, by LSBio (2 mentions)
Dynabeads sheep anti rat igg, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa, by BD (1 mentions)
Cd11b, by Thermo Fisher Scientific (1 mentions)
Facsaria iiu, by BD (1 mentions)
Igd 11 26c 2a, by Thermo Fisher Scientific (1 mentions)
Mac 1 m1 70, by Thermo Fisher Scientific (1 mentions)
Cd117 2b8, by Thermo Fisher Scientific (1 mentions)
Igm 11 41, by Thermo Fisher Scientific (1 mentions)
Facscalibur, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
Cd8α 53 6, by Thermo Fisher Scientific (1 mentions)
Cd45.1 a20, by BioLegend (1 mentions)
Cd45.2 104, by BioLegend (1 mentions)
4 protocols using tcrγδ gl3
1
Antigen-specific CD8+ T cell responses
2
Multiparameter Analysis of Hematopoietic Cells
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Bones, spleen, and thymus were dissected, crushed in PBS with 2% FCS and cells were collected after passing through a 70 μm filter. They were then Fc-blocked (CD16/32; 93) and stained with combinations of the antibodies Sca1 (D7), CD105 (MJ7/18), CD41 (MWReg30), CD48 (HM48-1), CD3 (145-2C11), CD4 (RM4-5), CD8 (53-6.7), B220 (RA3-6B2), NK1.1 (PK136), Mac1 (M1/70), Gr1 (RB6-8C5), TER119 (TER-119), CD150 (TCF15-12F12.2), CD117 (2B8, eBioscience), CD127 (A7R34), CD44 (IM7), CD25 (PC61.5, eBioscience), CD19 (1D3, eBioscience), TcRβ (H57-597, eBioscience), TcRγδ (GL3, eBioscience), Ly6C (AL-21), Ly6G (1A8), MHCII (M5/114.15.2), CD11c (N418), PDCA1 (927), Ly6D (49H4), Flt3 (A2F10), IgD (11-26c.2a), and IgM (11/41, eBioscience). All antibodies were purchased from BD Biosciences unless otherwise indicated. Propidium iodide (PI) was utilized to discriminate dead cells. For hematopoietic stem and progenitor cell isolation, cells were subjected to lineage depletion using Dynabeads sheep anti rat IgG (Life Technologies) together with TER119, CD19, CD3, Gr1, and CD11b antibodies prior to staining. Analysis and cell sorting was performed primarily on an LSR Fortessa and FACSAria IIu (BD Biosciences). Analysis of data was done using the Flowjo 9.9.6 software (Flowjo).
3
Antigen-specific CD8+ T cell responses
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Cx3cr1gfp/+ mice were infected with 2x108 Yptb-OVA, and on day 6 post infection LP cells were isolated from the ileum and cecum. Cells were stained with antibodies for CD11c, CD11b, I-Ab and dump gate (CD103, IgA (AD3, LSBio), CD19 (1D3, eBioscience), TCRβ (H57-597, eBioscience), TCRγδ (GL3, eBioscience)) sorted using a FACSAria (BD) to greater than 96% purity. Congenically marked CD8+CD44hi memory cells were sorted from Yptb-OVA memory mice and labelled with CFSE. 5x104 APCs and 5x104 memory T cells were mixed and incubated for 3 days with the addition of 5U/ml IL-2 for the final day of incubation. CFSE dilution was examined by flow cytometry and a live/dead stain was used to exclude dead cells from the analysis. Yptb- OVA memory cells were also incubated with APCs pulsed with SIINFEKL and YopE69-77 peptides to determine the maximum antigen specific response, and 60-70% of sorted Yptb-OVA memory cells divided in response to this peptide mixture.
4
Immunophenotyping of Intestinal Intraepithelial Lymphocytes
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For flow cytometry, IELs were stained with the indicated antibodies as previously described [25] . The following antibodies were used for flow cytometry: TCRβ (H57-597, eBioscience), TCRγδ (GL3, eBioscience), CD8α (53-6.7, eBioscience), CD8β (CT-CD8b, Invitrogen), CD4 (BD 552775), CD45.1 (A20, BioLegend), CD45.2 (104, BioLegend). FACS analysis was performed on a FACS Calibur flow cytometer quipped with Cell Quest software (BD Biosciences, Franklin Lakes, NJ). The gating strategy was shown in supplementary data as Figure S1.
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