Calcitriol (1,25(OH)2D3; Sigma-Aldrich Corp., St Louis, MO, USA) was solubilized in absolute ethanol to 1 mm and further diluted in X-VIVO 15 medium (04–418, Lonza, Basel, Switzerland) before addition to the cell culture. The antibodies used were fluorescein isothiocyanate-conjugated mouse anti-human CD4 (clone RTF-4 g, Southern Biotech, Birmingham, AL, USA), fluorescein isothiocyanate-conjugated mouse IgG1 isotype control (15H6, Southern Biotech), allophycocyanin-conjugated anti-human CD25 (IgG1, Immunotools, Friesoythe, Germany), allophycocyanin-conjugated mouse isotype IgG1 (Immunotools), mouse anti-VDR (D-6, sc-13133, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-GAPDH (6C5, sc-32233, Santa Cruz Biotechnology), rabbit anti-TAGAP (EPR15593, Abcam, Cambridge, UK), rabbit anti-Actin (A2066, Sigma-Aldrich Corp.), rabbit monoclonal IgG (EPR25A, Abcam), horseradish peroxidase-conjugated goat anti-mouse IgG and goat anti-rabbit IgG (Jackson ImmunoResearch Laboratories Europe Ltd., Suffolk, UK) and Alexa Fluor 647-conjugated goat anti-rabbit IgG (A-21245, Molecular Probes, Life Technologies, Carlsbad, CA, USA).
Fluorescein isothiocyanate conjugated mouse anti human cd4 clone rtf 4 g
Manufactured by Southern Biotech
Sourced in United States
Fluorescein isothiocyanate-conjugated mouse anti-human CD4 (clone RTF-4) is a monoclonal antibody that binds to the CD4 receptor on human T helper cells. It is conjugated with the fluorescent dye fluorescein isothiocyanate (FITC) for detection purposes.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 647 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Epr25a, by Abcam (1 mentions)
1 25 oh 2d3 calcitriol, by Merck Group (1 mentions)
X vivo 15 medium, by Lonza (1 mentions)
Attune acoustic focusing flow cytometer, by Thermo Fisher Scientific (1 mentions)
Mouse anti human cd8 clone hit8a, by BD (1 mentions)
Mouse igg1 isotype control 15h6, by Southern Biotech (1 mentions)
Easysep human cd8 positive selection kit, by STEMCELL (1 mentions)
2 protocols using fluorescein isothiocyanate conjugated mouse anti human cd4 clone rtf 4 g
1
Calcitriol Solubilization and Antibody Preparation
2
Isolation and Characterization of CD4+ and CD8+ T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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+ and CD8 + T cells CD4 + and CD8 + T cells were purified using immunomagnetic cell separation kits (Human CD4 + negative selection kit and EasySep™ Human CD8 + positive selection kit and EasySep™, STEMCELL Technologies, Vancouver, Canada) as described in (Rhead et al., 2018; Bos et al., 2015) . Purity of isolated T cells was confirmed to be at least 95% as measured by flow cytometry (Attune Acoustic Focusing Flow Cytometer, Life Technologies, Carlsbad, CA, USA). Antibodies used for purity assessment by flow cytometry were fluorescein isothiocyanateconjugated mouse anti-human CD4 (clone RTF-4 g, Southern Biotech, Birmingham, AL, USA), mouse anti-human CD8 (clone HIT8a, BD Biosciences, San Jose, CA, USA) and mouse IgG1 isotype control (15H6, Southern Biotech).
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