Spleens were isolated from tumor-naive female C57Bl/6N mice. After lysis of erythrocytes using the Mouse Erythrocyte Lysing Kit (R&D Systems) 2.5 × 105 splenocytes per well were then seeded in a 96-well round bottom plate and stimulated with 4 µg/ml plate-bound or 1 µg/ml soluble rat anti-mouse CD3 (clone 17A2; BD Pharmingen). Ferric chloride FeCl3 (Sigma Aldrich), ferric sulfate Fe2(SO4)3 (Sigma Aldrich), ferric citrate FeC6H5O7 (Sigma Aldrich), and holo-transferrin were added at concentrations of 2.5µM, 5 µM, 10 µM and 20 µM elementary iron. Splenocytes were cultured in RPMI-1640 medium (PAN Biotech) supplemented with 10% FCS (Biochrom), 2% sodium pyruvate (Sigma), 1× non-essential amino acids (Gibco), 0.01% β-mercaptoethanol (Roth), 1% penicillin/streptomycin (Lonza) and 2 mM L-glutamine (Lonza).
Soluble rat anti mouse cd3 clone 17a2
Manufactured by BD
Soluble rat anti-mouse CD3 (clone 17A2) is a laboratory reagent used in research applications. It is an antibody that binds to the CD3 receptor on mouse T cells. The product is provided in a soluble format.
Automatically generated - may contain errors
Lab products found in correlation
Fetal calf serum (fcs), by Harvard Bioscience (2 mentions)
Mouse erythrocyte lysing kit, by R&D Systems (2 mentions)
Ferric sulfate fe2 so4 3, by Merck Group (2 mentions)
Ferric citrate fec6h5o7, by Merck Group (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Ferric chloride fecl3, by Merck Group (2 mentions)
Sodium pyruvate, by Merck Group (2 mentions)
Penicillin streptomycin, by Lonza (2 mentions)
L glutamine, by Lonza (2 mentions)
Rpmi 1640 medium, by PAN Biotech (2 mentions)
Holo transferrin, by Merck Group (1 mentions)
β mercaptoethanol, by Lonza (1 mentions)
β mercaptoethanol, by Carl Roth (1 mentions)
2 protocols using soluble rat anti mouse cd3 clone 17a2
1
Iron Supplementation Impacts T Cell Activation
2
Investigating Iron-Induced T Cell Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Spleens were isolated and after lysis of erythrocytes using the Mouse Erythrocyte Lysing Kit (R&D Systems) 2.5 x 105 splenocytes per well were then seeded in a 96-well round bottom plate and stimulated with 4 µg/ml plate-bound or 1 µg/ml soluble rat anti-mouse CD3 (clone 17A2; BD Pharmingen). Ferric chloride FeCl3 (Sigma Aldrich), ferric sulfate Fe2(SO4)3 (Sigma Aldrich), ferric citrate FeC6H5O7 (Sigma Aldrich) were added at concentrations of 2.5, 5, 10 and 20 µM elementary iron. Splenocytes were cultured in RPMI-1640 medium (PAN Biotech) supplemented with 10% FCS (Biochrom), 2% sodium pyruvate (Sigma), 1× non-essential amino acids (Gibco), 0.01% β-mercaptoethanol (Roth), 1% penicillin/streptomycin (Lonza) and 2 mM L-glutamine (Lonza).
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