TNFR1 Antibody / TNFRSF1A Polyclonal Antibody (R32696)

TNFR1 Antibody WB in HeLa Wild-Type and TNFRSF1A Knockout Cells. Western blot analysis of human HeLa wild-type (lane 1) and TNFRSF1A knockout (lane 2) whole-cell lysates using the TNFRSF1A Polyclonal Antibody. A prominent band at approximately 60-65 kDa is detected in wild-type cells and is substantially reduced in the knockout sample, with residual signal at the corresponding position. The predicted molecular weight of TNFR1 is approximately 50 kDa, although glycosylation can increase its apparent molecular weight during SDS-PAGE. Samples were loaded at 30 ug per lane and separated by 10% SDS-PAGE under reducing conditions. The primary antibody was used at 0.5 ug/ml, with GAPDH serving as the loading control.

TNFR1 Antibody WB in Human Cell Lines and Rat and Mouse Tissues. Western blot analysis of human HeLa (lane 1), A549 (lane 2), and Raji (lane 3) whole-cell lysates; rat brain (lane 4) and lung (lane 5); and mouse brain (lane 6) and lung (lane 7) tissue lysates using the TNFRSF1A Polyclonal Antibody. Immunoreactive bands are detected at approximately 55-70 kDa across the seven samples, compared with the predicted molecular weight of approximately 50 kDa for TNFR1. The higher apparent molecular weights may reflect glycosylation of TNFR1, although the modifications responsible for individual bands have not been confirmed. Multiple bands are particularly evident in A549 cells, and the identity of these additional species has not been established. Samples were loaded at 30 ug per lane and separated by 10% SDS-PAGE under reducing conditions. The primary antibody was used at 0.5 ug/ml.

TNFR1 Antibody IF in Human A549 Cells. Immunofluorescence staining of human A549 cells using the TNFRSF1A Polyclonal Antibody at 5 ug/ml, detected with Fluoro488-conjugated goat anti-rabbit IgG (green), with DAPI nuclear counterstaining (blue). TNFR1 immunoreactivity is observed predominantly in the cytoplasm, exhibiting a granular and punctate distribution with areas of perinuclear enrichment. Staining is also visible along portions of the cell periphery, while the nuclei show comparatively limited immunoreactivity. Enzymatic antigen retrieval was performed for 15 minutes, followed by blocking with 10% goat serum. The primary antibody was incubated overnight at 4°C.

TNFR1 Antibody FACS in Human U937 Cells. Flow cytometry analysis of human cells using the TNFRSF1A Polyclonal Antibody at 1 ug per million cells, detected with DyLight 488-conjugated goat anti-rabbit IgG. The antibody-stained population (blue) exhibits a clear rightward fluorescence shift relative to the rabbit IgG isotype control (green) and unlabeled cells (red), demonstrating TNFR1-associated immunoreactivity. Cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. The primary antibody was incubated for 30 minutes at 20°C, followed by secondary antibody incubation for 30 minutes at 20°C.