human immunoglobulin a2  (Elabscience Biotechnology)

Journal: Biomedicines

Article Title: Stearic Acid and TNF-α Co-Operatively Potentiate MIP-1α Production in Monocytic Cells via MyD88 Independent TLR4/TBK/IRF3 Signaling Pathway

doi: 10.3390/biomedicines8100403

Figure Lengend Snippet: Disruption of TLR4 suppresses cooperative effect of stearic acid with TNF-α for MIP-1α/CCL3 production. ( A , B ) THP-1 cells were incubated with oxidized 1-palmitoyl-2-arachidonyl-sn- glycero-3-phosphorylcholine (OXPAPC) (30 µg/mL) for 1 h and then treated with stearic acid and TNF-α for 24 h. Cells and culture media were collected. MIP-1α/CCL3 gene expression was determined by real time PCR and secreted MIP-1α/CCL3 protein was determined in culture media by ELISA. ( C , D ). Monocytic cells were treated with 2 µg/mL of neutralizing TLR4 mAb or isotype-matched control (IgA2) for 40 min. Antibody-treated cells were treated with stearic acid and TNF-α for 24 h. MIP-1α/CCL3 mRNA and secreted MIP-1α/CCL3 protein were determined. ( E – G ) Monocytic cells were transfected with either control or TLR4 siRNA. TLR4-deficient cells were stimulated with stearic acid and TNF-α for 24 h. Knockdown efficiency of transfection was checked by TLR4 gene expression with qRT-PCR. MIP-1α/CCL3 mRNA and secreted MIP-1α/CCL3 protein were determined. The results obtained from minimum three independent experiments with three replicates of each experiment are shown. All data are expressed as mean ± SEM ( n ≥ 3); * P < 0.05; ** P < 0.01when compared with Stearic acid or TNF-α alone.

Article Snippet: TLR4 neutralizing antibody (cat#: mabg-htlr4) and Immunoglobulin A2 (IgA2) isotype control (cat#: maba2-ctrl) were purchased from InvivoGen (San Diego, CA, USA).

Techniques: Disruption, Incubation, Gene Expression, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Transfection, Knockdown, Quantitative RT-PCR

Journal: Frontiers in Medicine

Article Title: Predicting cardiometabolic disease in medical students using FibroScan and 30-year Framingham risk scores

doi: 10.3389/fmed.2024.1431935

Figure Lengend Snippet: Comparison of participants with and without elevated liver stiffness.

Article Snippet: To measure human immunoglobulin A2 and human PIGR levels, we used a commercially available enzyme-linked immunosorbent assay kit from Elabscience Biotechnology Inc., United States.

Techniques: Comparison, Control, Clinical Proteomics

Journal: Frontiers in Microbiology

Article Title: S-Layer Protein of Lactobacillus helveticus SBT2171 Promotes Human β-Defensin 2 Expression via TLR2–JNK Signaling

doi: 10.3389/fmicb.2019.02414

Figure Lengend Snippet: Mechanisms of hBD2 upregulation by LH2171 SLP. Caco-2 cells were pre-incubated with anti-TLR2 antibody (a-TLR2), isotype control antibody (IgA2) (A,C,D) , or the indicated inhibitor (SB202190 for p38, SP600125 for JNK, PD98059 for ERK, and JSH-23 for NF-κB) (B) and then treated with LH2171 SLP. hBD2 mRNA levels were evaluated by quantitative real time-PCR (A,B) , and protein levels of total JNK and phospho-JNK (P-JNK) (C) or total c-Jun and phospho-c-Jun (D) in total cell lysates were analyzed by western blotting. Each experiment was performed in triplicate; data are shown as mean ± SD (A–D) . ∗∗ P < 0.01 and ∗∗∗ P < 0.001 vs. IgA2 + SLP (A) or SLP (B) (one-way ANOVA and Dunnett’s post hoc test). Values not sharing a common letter are significantly different ( P < 0.05 by one-way ANOVA and Tukey–Kramer post hoc test) (C,D) .

Article Snippet: The next day, the medium was changed to DMEM without serum and antibiotics followed by incubation for 24 h. The cells were then treated with anti-TLR2 antibody (Invivogen, San Diego, CA, United States) for 1 h. In accordance with the manufacturer’s instructions, a human immunoglobulin A2 (IgA2) isotype (Invivogen) was used as a control to prevent non-specific binding and blocking activities of the antibody.

Techniques: Incubation, Real-time Polymerase Chain Reaction, Western Blot