CD3+ T cells were isolated from PBMC as above and incubated for 24h either unstimulated or with 1μg/ml CD3 and CD28 antibodies (BD Biosciences), together with 10ng/ml IL-1β or 100ng/ml truncated IL-36α, β or γ. Cultures were subsequently prepared for qRT-PCR or flow cytometry. Cells were washed in FACS buffer (PBS+0.5%BSA+0.1%NaN3) then stained with antibodies against CD3 (clone S4.1, Invitrogen), CD4 (OKT-4, eBioscience), CD8 (SK1, BD), CLA (HECA-452, Biolegend), CD103 (Ber-ACT8, Biolegend), CD25 (BC96, Biolegend), CD69 (FN50, Biolegend), CD54 (HCD54, Biolegend) and appropriate isotype-matched control antibodies for 30 min at 4°C in the dark. After 2 washes in FACS buffer, cells were analyzed using a BD LSR2 flow cytometer gating on lymphocytes expressing CD3 and CD4 or CD8.
Cla heca 452
Manufactured by BioLegend
The CLA (HECA-452) is a lab equipment product that serves as a cell surface marker. It is used for the identification and enumeration of cells expressing the CLA (Cutaneous Lymphocyte Antigen) protein.
Automatically generated - may contain errors
Lab products found in correlation
Lsr 2 flow cytometer, by BD (1 mentions)
Cd8 sk1, by BD (1 mentions)
Cd103 ber act8, by BioLegend (1 mentions)
Cd69 fn50, by BioLegend (1 mentions)
Cd25 bc96, by BioLegend (1 mentions)
Cd4 okt4, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Recombinant human pdgf, by R&D Systems (1 mentions)
Streptavidin pe, by BioLegend (1 mentions)
Hcc1806, by American Type Culture Collection (1 mentions)
Cd44 fitc im7, by BioLegend (1 mentions)
Neuraminidase s, by New England Biolabs (1 mentions)
Pngase f, by New England Biolabs (1 mentions)
2 protocols using cla heca 452
1
T cell activation and phenotyping
2
Characterization and Purification of Glycoproteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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SLBR-N, SLBR-H, and SK678 were characterized and purified as previously described (15 (link)). Neuraminidase S and PNGaseF were purchased from New England Biolabs (NEB), and recombinant human PDGF was purchased from R&D Systems. Flow cytometry antibodies and lectins were acquired as follows: integrin β4 (439-9b; Abcam, ab110167), CD24–allophycocyanin (APC; ML5, BioLegend), CD24–phycoerythrin (PE; ML5, BioLegend), CD44-FITC (IM7, BioLegend), SNA (B-1305-2, Vector Laboratories), PE streptavidin (405203, BioLegend), CLA (HECA-452, BioLegend), anti-rat immunoglobulin M (IgM) FITC (MRM-47, BioLegend), anti–glutathione S-transferase (GST) APC (Columbia Biosciences), HA-FITC (385906, EMD Millipore), and anti–CA19-9 (121SLE, Novus Biologicals). Immunoblotting antibodies were acquired as follows: tubulin [Cell Signaling Technology (CST), 3873], CD44 (Abcam, 157107), FUT3 (Abcam, 110082), anti-rat IgM horseradish peroxidase (SouthernBiotech), phospho-PDGFRβ Tyr1009 (CST, 3124), PDGFRβ (CST, 3169), phospho-STAT3 Tyr705 (CST, 9145), and STAT3 (CST, 9139).
HMLER cells were provided by R. Weinberg (Massachusetts Institute of Technology). Cells were cultured in MEBM mammary epithelial cell growth basal medium (Lonza). MDA-MB-231-TE3 was provided by S. Tavazoie (Rockefeller University). MDA-MB-231 and HCC1806 were obtained from the American Type Culture Collection.
HMLER cells were provided by R. Weinberg (Massachusetts Institute of Technology). Cells were cultured in MEBM mammary epithelial cell growth basal medium (Lonza). MDA-MB-231-TE3 was provided by S. Tavazoie (Rockefeller University). MDA-MB-231 and HCC1806 were obtained from the American Type Culture Collection.
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