negative selection magnetic beads  (Miltenyi Biotec)

Journal: mAbs

Article Title: Higher order receptor clustering due to the IgG3 subclass is necessary for TLR4 signaling and tolerance induction by novel human anti-TLR4 antibodies

doi: 10.1080/19420862.2025.2515415

Figure Lengend Snippet: IgG3 TV-7D11 induces tolerance in primary human myeloid cells. Human CD14+ monocytes were purified by magnetic bead negative selection. 1 × 10^6 cells/well were seeded in 24-well plates and stimulated with IgG3 8H1 (10 μg/ml), IgG3 TV-7D11 (10 μg/ml), or LPS (.01 μg/ml). After 24 h supernatants were removed and tested for TNF-α by ELISA (a), followed by addition of fresh media and culture for 5 d to allow cells to return to a resting state (b). On day 6, cells were split into 96-well plates at 50,000 cells/well and allowed to rest for 24 h prior to re-challenge with LPS at .01 μg/ml. Supernatants were again collected at 24 h and assayed for TNF-α production by ELISA (c). Data points represent three individuals from two independent experiments. Statistical analysis by ANOVA * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human CD14+ monocytes were purified by magnetic bead negative selection (Miltenyi, #130–117–337).

Techniques: Purification, Selection, Enzyme-linked Immunosorbent Assay