A single-cell suspension from mouse spleen was prepared using gentleMACS Dissociator (no. 130-093-235; Miltenyi Biotec). CD4+ and CD8+ T cells were then isolated from this single-cell suspension using the magnetic CD4+ T Cell Isolation Kit (no. 130-104-454; Miltenyi Biotec) or CD8+ T Cell Isolation Kit (no. 130-104-075; Miltenyi Biotec) according to the manufacturer’s instructions.
Magnetic cd4 t cell isolation kit
Manufactured by Miltenyi Biotec
The Magnetic CD4+ T cell isolation kit is a lab equipment product from Miltenyi Biotec designed for the isolation of CD4+ T cells from various cell samples. The kit utilizes magnetic separation technology to selectively enrich the target cell population.
Automatically generated - may contain errors
Lab products found in correlation
Cd8 t cell isolation kit, by Miltenyi Biotec (2 mentions)
Gentlemacs dissociator, by Miltenyi Biotec (2 mentions)
Fitc or percp cy5.5 anti cd44 clone im7, by BioLegend (2 mentions)
Moflo facs sorter, by Agilent Technologies (2 mentions)
7 aad, by Thermo Fisher Scientific (2 mentions)
Facsaria 3 sorter, by BD (2 mentions)
Charcoal stripped fbs, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 800, by Thermo Fisher Scientific (1 mentions)
Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 680, by Thermo Fisher Scientific (1 mentions)
Imagelite software, by LI COR (1 mentions)
Ccl19, by R&D Systems (1 mentions)
Odyssey buffer, by LI COR (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
6 protocols using magnetic cd4 t cell isolation kit
1
Isolation of Mouse CD4+ and CD8+ T Cells
2
Isolation of Mouse CD4+ and CD8+ T Cells
3
Chemotaxis Assay for Thymocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD4 SP thymocytes from WT and X-MAID mice were isolated by negative selection using a magnetic CD4+ T cell isolation kit (Miltenyi) and resuspended in DMEM with 10% charcoal stripped FBS (Gibco). 2x105 cells per well were placed in the top chambers of a 24 well transwell plate (5μm pore size, Corning) and allowed to settle at 37°C for 10 minutes. The top chambers were then placed on top of bottom chambers containing 100ng/ml CCL19, and incubated for 2 hours at 37°C. Top chambers were then removed and cells in the bottom chambers were counted using a hemocytometer. Percent migration was calculated based on input cell numbers.
4
CD4+ T Cell Subset Isolation
5
CCL19-Induced Signaling in Thymocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD4 SP thymocytes from WT and X-MAID mice were isolated by negative selection using a magnetic CD4+ T cell isolation kit (Miltenyi). Cells were starved for 4 hours in serum-free DMEM (Corning) before stimulation 100ng/ml CCL19 (R&D Systems) for the indicated times. Cells were then lysed with 1% Triton X100, 50 mM Tris-HCl, 50 mM NaCl, 5 mM EDTA, 50 mM NaF, 30 mM Na4P2O7, 50 mM β-glycerophosphate, with Roche Complete protease inhibitors. Proteins were separated by SDS/PAGE (4-12% NuPAGE bis-tris gel, Invitrogen) transferred to nitrocellulose membranes, blocked with Odyssey buffer (Licor) and probed for the indicated proteins. Antibodies to pERK1/2 (Thr202/Tyr204) and pAkt (Ser473) were from Cell Signaling Technologies. GAPDH was used as a loading control, and was detected with Clone 6C5 antibody, from Sigma. Primary antibodies were detected with fluorescent secondary antibodies (anti-rabbit AlexaFluor 800 or anti-mouse AlexaFluor 680, from Invitrogen). Blots were imaged using an Odyssey fluorescence-based imaging system (Licor) and prepared for publication using ImageLite Software (Licor).
6
Isolation and Sorting of CD4+ T Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Retina and secondary lymphoid organs (SLO, including draining LN and spleens) from CAU mice (week 12 -16 post-induction) were harvested and pooled, and CD4 + T cells were isolated via negative selection with a magnetic CD4 + T cell isolation kit (Cat #: 130-104-454, Miltenyi Biotec Inc.). The isolated cells were >98% CD4 + as confirmed by flow cytometry, and they were stained with FITC-or PerCP/Cy5.5-anti-CD44 (clone IM7, Cat #: 103006 or 103032, BioLegend) antibody, and sorted for CD44 hi memory and CD44 -~lo control subpopulations using a MoFlo ® FACS sorter (Dako Cytomation) or a BD FACSAria ™ III sorter (BD Biosciences). The sorted cells in equal live numbers (1×10 5 ) were cultured for 5 days and stained with 7-AAD (Cat #: 00-6993-50, ThermoFisher), and the viable cells were determined as 7-AAD unstained populations by flow cytometric analysis.
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!