Porcine TNF-alpha Antibody

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Porcine TNF‑ alpha Antibody. Recombinant Porcine TNF-a (Catalog # 690-PT) induces cytotoxicity in the the PK-15 porcine kidney epithelial cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Cytotoxicity elicited by Recombinant Porcine TNF-a (0.05 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Porcine TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF690). The ND₅₀ is typically 0.2-0.8 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

TNF‑ alpha in Porcine PBMCs. TNF-a was detected in immersion fixed porcine peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA using Goat Anti-Porcine TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF690) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Porcine TNF-alpha by Western Blot Inflammatory response in PAMs induced by PRRSV 5′UTR RNA and LPS. A, B PAMs were transfected with different doses of 5′UTR RNA (1, 2, and 4 μg/well) along with 1 μg/mL LPS. qRT-PCR and Western blot results showed that compared with PAMs in other groups, PAMs in the 4 μg 5′UTR RNA and 1 μg/mL LPS co-stimulation group produced higher levels of IL-1 beta (p < 0.05). C After co-stimulation, the relative expression level of IL-1 beta mRNA obtained was similar at the 12 and 24 h time-points. D 5′UTR RNA and LPS co-stimulation induced IL-1 beta expression in cells and supernatants. E, F 5′UTR RNA and LPS co-stimulation induced increased levels of IL-6, IL-8 and TNF-alpha mRNA and protein. Expression was normalized to that of GAPDH. Different letters (a, b, c, d, and e) on data indicate significant differences between groups (p < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31300043), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Porcine TNF-alpha by Western Blot Inflammatory response in PAMs induced by PRRSV 5′UTR RNA and LPS. A, B PAMs were transfected with different doses of 5′UTR RNA (1, 2, and 4 μg/well) along with 1 μg/mL LPS. qRT-PCR and Western blot results showed that compared with PAMs in other groups, PAMs in the 4 μg 5′UTR RNA and 1 μg/mL LPS co-stimulation group produced higher levels of IL-1 beta (p < 0.05). C After co-stimulation, the relative expression level of IL-1 beta mRNA obtained was similar at the 12 and 24 h time-points. D 5′UTR RNA and LPS co-stimulation induced IL-1 beta expression in cells and supernatants. E, F 5′UTR RNA and LPS co-stimulation induced increased levels of IL-6, IL-8 and TNF-alpha mRNA and protein. Expression was normalized to that of GAPDH. Different letters (a, b, c, d, and e) on data indicate significant differences between groups (p < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31300043), licensed under a CC-BY license. Not internally tested by R&D Systems.