mouse anti tlr2 ab  (Hycult Biotech)

Journal: European journal of immunology

Article Title: TLR2 on CD4+ and CD8+ T cells promotes control of Mycobacterium tuberculosis infection.

doi: 10.1002/eji.202350715

Figure Lengend Snippet: Figure 1. Deletion of exon 3 of Tlr2 in CD4+ and CD8+ T cells in Tlr2fl/flxCd4cre/cre mice. (A) Location of LoxP sites around exon 3 of Tlr2 in Tlr2fl/fl

Article Snippet: Blots were either incubated with polyclonal goat anti-mouse TLR2 Ab (R&D Systems; AF1530) or β-actin (R&D Systems; MAB8929), followed by HRP-conjugated donkey anti-goat IgG(H+L) mAb (Jackson) and bands detected by chemiluminescence using Pierce ECL Plus substrate (Fisher).

Techniques:

Journal: Journal of Innate Immunity

Article Title: HMGB1 Binds to Lipoteichoic Acid and Enhances TNF-a and IL-6 Production through HMGB1-Mediated Transfer of Lipoteichoic Acid to CD14 and TLR2

doi: 10.1159/000369972

Figure Lengend Snippet: HMGB1 interacts with LTA and mediates the transfer of LTA to sCD14. a Native PAGE mobility shift assay of the transfer of LTA to sCD14 following HMGB1-LTA binding. A mixture of sLTA and sCD14 was incubated with HMGB1 for 12 h at 37°C and then subjected to native PAGE analysis on a 4–15% gradient gel, with subsequent silver staining (upper panel). LBP was used as a positive control. Lower panel represents SDS-PAGE analysis as a protein loading control. The position of uncomplexed sCD14 and HMGB1 is indicated by the asterisk. b Western blot analysis using an anti-CD14 Ab after native PAGE. Asterisk indicates uncomplexed sCD14. c HMGB1-mediated transfer of BODIPY FL-LTA to CD14. A mixture of BODIPY FL-sLTA and sCD14 was incubated in the presence or absence of HMGB1 for 10 h at 25°C and the fluorescence level was measured. LBP was used as a positive control, and 2% SDS was used to solubilize the disaggregated BODIPY FL-sLTA completely to maximize the fluorescence. * p < 0.05, ** p < 0.01 (n = 4). d Transfer of BODIPY FL-sLTA to TLR2. BO = BODIPY FL. HEK293-hTLR2/6 cells were stimulated with a mixture of 1 μg/ml of BODIPY FL-sLTA and 0.5 μg/ml of sCD14 in the presence or absence of 1 μg/ml of HMGB1 for 4 h. LBP (1 μg/ml) was used as a positive control. The cells were then fixed and stained with an anti-TLR2 Ab. Scale bar = 10 μm.

Article Snippet: The cells were washed with PBS, blocked with 1% BSA in PBS for 10 min and incubated with mouse anti-TLR2 Ab (eBioscience) overnight at 4°C.

Techniques: Clear Native PAGE, Mobility Shift, Binding Assay, Incubation, Silver Staining, Positive Control, SDS Page, Control, Western Blot, Fluorescence, Staining