negative beads selection kit  (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

Journal: Cell Death Discovery

Article Title: CXCL16 knockout inhibit asthma airway inflammation by suppressing H2-DM molecular mediated antigen presentation

doi: 10.1038/s41420-025-02371-6

Figure Lengend Snippet: A GO enrichment of biological process for downregulated differentially expressed genes in CXCL16-knockout mouse BMDCs versus wild-type mouse BMDCs after Aspergillus challenge ( n = 3). B CD11c + BMDCs from wild-type mice or CXCL16-knockout mice were pulsed with 20 μg OVA 323-339 peptide and incubated at a 1:10 ratio with CFSE-labeled splenic CD4 + OT-II T cells for 4 days. OVA-specific proliferation is presented as a proliferation index ( n = 4). C Phagocytosis of FITC-dextran by CD11c + sorting BMDCs in vitro ( n = 3). D Unsupervised clustering of genes involved in biological processes of antigen processing and presentation, MHC class II biosynthetic process, regulation of phagocytosis, phagocytosis, and recognition were analyzed ( n = 3). E Real-time PCR was performed to detect downregulated differentially expressed PRR genes of Aspergillus -challenged CXCL16-knockout BMDCs and wild-type BMDCs ( n = 3). F The mRNA level of H2-DM on Aspergillus -induced CXCL16-knockout mouse BMDCs or wild-type mouse BMDCs ( n = 6). G The mRNA level of H2-Oa and H2-Ob in CXCL16-knockout BMDCs ( n = 6). * P < 0.05, ** P < 0.01 Aspergillus -challenged CXCL16-knockout mice versus wild-type mice. A.f . stands for Aspergillus .

Article Snippet: Naïve CD4 + T cells were purified from the spleens of OT-II mice by a CD4 + bead negative selection kit (MACS Miltenyi Biotec) and were labeled with 5 μM CFSE (ThermoFisher Scientific) at 37 °C for 30 min.

Techniques: Knock-Out, Incubation, Labeling, In Vitro, Real-time Polymerase Chain Reaction