SAPK4 Antibody / Stress-activated protein kinase 4 / MAPK13 (R31807)

Immunofluorescence analysis of SAPK4 in human U2OS cells. SAPK4 antibody was used to detect SAPK4 (green) in human U2OS cells, with alpha tubulin antibody labeling the cytoskeleton (red). Cells were enzymatically treated for antigen retrieval, blocked with goat serum, and incubated with primary antibodies overnight. Secondary antibodies used were Fluoro488-conjugated anti-rabbit IgG and Cy3-conjugated anti-mouse IgG. SAPK4 signal is observed in the cytoplasmic compartment.

Immunohistochemistry analysis of SAPK4 in human breast cancer tissue. SAPK4 antibody was used to detect SAPK4 in paraffin-embedded human breast cancer tissue. Antigen retrieval was performed using EDTA buffer (pH 8.0). Tumor epithelial cells show cytoplasmic staining, while surrounding stromal cells display weaker signal. Detection was performed using an HRP-DAB system.

Immunohistochemistry analysis of SAPK4 in human ovary cancer tissue. SAPK4 antibody was applied to paraffin-embedded human ovary cancer tissue following EDTA-based antigen retrieval (pH 8.0). Cytoplasmic staining is observed in tumor epithelial cells with lower background in adjacent stromal regions. Signal was visualized using HRP-DAB chemistry.

Immunohistochemistry analysis of SAPK4 in mouse colon tissue. SAPK4 antibody was used to stain paraffin-embedded mouse colon tissue after EDTA antigen retrieval (pH 8.0). Positive cytoplasmic staining is present in epithelial cells lining the intestinal crypts, with minimal staining in surrounding connective tissue.

Immunohistochemistry analysis of SAPK4 in rat colon tissue. Paraffin-embedded rat colon tissue was stained using SAPK4 antibody following EDTA-mediated antigen retrieval (pH 8.0). SAPK4 signal is observed primarily in epithelial cells of the intestinal mucosa, with lower signal in non-epithelial compartments.

Western blot analysis of SAPK4 using anti-SAPK4 antibody. Proteins from human RT4, TE-1, A431, and Caco-2 whole cell lysates (lanes 1-4), rat small intestine tissue lysate (lane 5), and mouse small intestine tissue lysate (lane 6) were separated by 10% SDS-PAGE under reducing conditions and transferred to a nitrocellulose membrane. The membrane was probed with rabbit anti-SAPK4 antibody followed by HRP-conjugated goat anti-rabbit IgG, and signal was detected by ECL. A specific band corresponding to SAPK4 was observed at approximately 42 kDa, consistent with the predicted molecular weight of SAPK4. In mouse small intestine, additional lower molecular weight bands were detected, which may reflect proteolytic processing, tissue-specific degradation, or partial cleavage products commonly observed in intestinal lysates with high protease activity.

Immunoprecipitation-western blot analysis of SAPK4 in human RT4 whole cell lysate using anti-SAPK4 antibody. RT4 whole cell lysate was subjected to immunoprecipitation followed by western blot detection using anti-SAPK4 antibody. Lane 1 shows human RT4 whole cell lysates (30 ug). Lane 2 shows rabbit control IgG used instead of anti-SAPK4 antibody in RT4 whole cell lysate. Lane 3 shows anti-SAPK4 antibody (2 ug) incubated with RT4 whole cell lysate (500 ug). A specific band corresponding to SAPK4 is detected at approximately 42 kDa, consistent with the predicted molecular weight of SAPK4. Additional lower molecular weight bands observed in the immunoprecipitated sample are consistent with residual IgG light chain signal and possible proteolytic fragments generated during immunoprecipitation and sample processing, which are commonly observed in IP-western blot analyses.

Flow cytometry analysis of fixed and permeabilized human Caco-2 cells with SAPK4 antibody at 1ug/million cells (blocked with goat sera); Red=cells alone, Green=isotype control, Blue= SAPK4 antibody.