In order to analyze immune cells and their anti-tumoral activity, B6 mice were challenged and swarm at TT and BT (n=9 per group) and then 3 mice of them were randomly sacrificed every wk. Primary cells were collected from lymph node (LN), draining lymph node (dLN), and spleen, and analyzed via FACS Canto II or FACS Aria I. Data were analyzed using FlowJo version 10 (TreeStar, San Carlos, CA, USA). Antibodies with the following specificities were used for staining: CD8α (53-6.7), IFNγ (XMG1.2), IL-4 (11B11), CD44 (IM7), CD62L (MEL-14), NK1.1 (PK136), TCRβ (H57-597), and γδTCR (GL3) from BioLegend (San Jose, CA, USA); CD4 (GK1.5) from Thermo Fisher Scientific (Waltham, MA, USA); and IL-17 (TC11-18H10) from BD Biosciences (San Jose, CA, USA). Anti-mouse CD16/32 (2.4G2; BioLegend) blocked the non-specific binding of the antibodies. For intracellular cytokine staining, the cells were stimulated with PMA (Merck Millipore, Burlington, MA, USA) and ionomycin (Santa Cruz, Dallas, TX, USA) in the presence of brefeldin A (Thermo Fisher Scientific), which were then fixed and permeabilized with an IC fixation buffer (Thermo Fisher Scientific).
Cd4 gk1.5
Manufactured by Thermo Fisher Scientific
Sourced in United States
The CD4 (GK1.5) is a lab equipment product. It is a monoclonal antibody that recognizes the CD4 antigen, which is expressed on the surface of T helper cells. This product can be used for various research applications involving the identification and analysis of CD4+ T cells.
Automatically generated - may contain errors
Lab products found in correlation
Facs aria 1, by Tree Star (2 mentions)
Anti mouse cd16 32 2.4g2, by BioLegend (2 mentions)
Fixable viability dye, by Thermo Fisher Scientific (2 mentions)
Cd90.1 ox 7, by Thermo Fisher Scientific (2 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Ic fixation buffer, by Thermo Fisher Scientific (1 mentions)
γδtcr gl3, by BioLegend (1 mentions)
Il 17 tc11 18h10, by BD (1 mentions)
Ionomycin, by Santa Cruz Biotechnology (1 mentions)
Facscanto 2, by Tree Star (1 mentions)
Facscanto 2, by BD (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Cd11b, by Thermo Fisher Scientific (1 mentions)
B220 ra3 6b2, by Thermo Fisher Scientific (1 mentions)
Ly6g 1a8, by BioLegend (1 mentions)
Ly6c hk1, by BioLegend (1 mentions)
Cd8a 53 6.7, by Thermo Fisher Scientific (1 mentions)
70 μm cell strainer, by Corning (1 mentions)
Anti mouse cd16 32, by BioLegend (1 mentions)
5 protocols using cd4 gk1.5
1
Comprehensive Immune Cell Analysis
2
Tracking T-cell Responses in Skin Inflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
These experiments were performed as previously described [14 (link),15 (link)] with slight modifications. Briefly, mice received intravenous (i.v.) transfer of CFSE-labeled, congenically marked 5x105 TEα cells 1 day before or 13 days after the intradermal exposure to mRNA(eGFP)-LNP or PBS. One μg of anti-Langerin-Eα or PBS were administered i.v. on day 14. Mice were then sacrificed 4 days later and the skin draining lymph nodes (axillary and brachial) harvested. Single-cell suspensions were stained with fixable viability dye (Thermo Fisher), CD4 (GK1.5), and CD90.1 (OX-7). The antibodies were purchased from BioLegend.
3
Antigen-specific CD4+ T cell response analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
These experiments were performed as previously described [14 (link),15 (link)] with slight modifications. Briefly, mice received intravenous (i.v.) transfer of CFSE-labeled, congenically marked 5×105 TEα cells 1 day before or 13 days after the intradermal exposure to mRNA(eGFP)-LNP or PBS. One μg of anti-Langerin-Eα or PBS were administered i.v. on day 14. Mice were then sacrificed 4 days later and the skin draining lymph nodes (axillary and brachial) harvested. Single-cell suspensions were stained with fixable viability dye (Thermo Fisher), CD4 (GK1.5), and CD90.1 (OX-7). The antibodies were purchased from BioLegend.
4
Tumor and Splenic Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tumour flow cytometry-based immuno phenotype was performed according to already published protocols45 (link). Briefly, tumours were minced and digested for about 1 hr at 37 °C under continuous mixing with a digestive mix containing 1 mg/mL collagenase IV, 0.1 mg/mL hyaluronidase, and 30 U/mL DNAse in RPMI 1640, all purchased from Sigma-Aldrich. The cell suspension was separated from the undigested material using a 70-μm cell strainer (Corning). On the contrary, the analysis of circulating leukocytes was performed using splenocytes collected by mechanical disruption of the tissue followed by red blood cell lysis with the ACK buffer (Lonza). One million of cells were incubated with anti-mouse CD16/32 (Biolegend) and subsequently stained with the following antibodies according to the vendor’s instructions: CD3 (17A2, Thermo Fisher Scientific), CD4 (GK1.5, Thermo Fisher Scientific), CD8a (53.6.7, Thermo Fisher Scientific), F4/80 (A3-1, Bio-Rad), MHC2 (M5/114.15.2, Thermo Fisher Scientific), CD11b (M1/70, Thermo Fisher Scientific), B220 (RA3-6B2, Thermo Fisher Scientific) CD11c (N418, Thermo Fisher Scientific), LY6C (HK1.4, Biolegend), LY6G (1A8, Biolegend), CD45 (30F11, Biolegend), Samples were acquired on a FACS Canto II (BD Biosciences) and analyzed with FlowJo software (FlowJo LLC).
5
Flow Cytometry Analysis of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were collected from LN and SP, and analyzed via FACS Canto or FACS Aria I. The data were analyzed using FlowJo version 10 (Version 10, Tree Star, Ashland, OR, USA). Antibodies with the following specificities were used for staining: CD8α (53–6.7), CD44 (IM7), NK1.1 (PK136), TCRβ (H57–597), γδTCR (GL3), and CD122 (IL-2R), all from BioLegend (San Jose, CA, USA); B220 (RA3–6B2) from BD Biosciences (San Jose, CA, USA); CD4 (GK1.5) from Thermo Fisher (Waltham, MA, USA), fluorochrome-conjugated CD1d tetramers loaded with PBS-567 and unloaded controls were obtained from the NIH tetramer facility (Emory University, Atlanta, GA, USA). Antimouse CD16/32 (2.4G2; BioLegend, San Jose, CA, USA) blocks nonspecific binding of antibodies.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!