proteome profiler human phospho mapk array membranes  (R&D Systems)

Journal: Scientific reports

Article Title: STEAP2 promotes hepatocellular carcinoma progression via increased copper levels and stress-activated MAP kinase activity.

doi: 10.1038/s41598-024-63368-2

Figure Lengend Snippet: Figure 4. STEAP2 Knockdown Decreases JNK and p38 phosphorylation. (a) The results of the phospho- MAPK protein array analysis using STEAP2 knockdown and control cells. Cell lysates were assessed using the Proteome Profiler Human Phospho-MAPK Array membranes (ARY003B, R&D Systems) overnight at 4 °C. Biotinylated detection antibodies were applied, and membranes were visualized using chemiluminescence. (b) The graph shows the pixel intensity of proteins with significant difference in STEAP2 knockdown (KD) cells compared to matched control cells. Densitometry analysis was performed using the Protein Array Analyzer for ImageJ. The decreased phosphorylation patterns of JNK isoforms at T183/Y185 (c) and p38 isoforms at T180/Y182 (d) due to STEAP2 KD were validated by Western blot. Western blot image was cropped to show appropriate protein band (SNU398 blot cropped). GAPDH protein level was used to validate equal sample loading.

Article Snippet: Cell lysates were assessed using the Proteome Profiler Human Phospho-MAPK Array membranes (ARY003B, R&D Systems) overnight at 4 °C.

Techniques: Knockdown, Phospho-proteomics, Protein Array, Control, Western Blot

Journal: Cancer Cell International

Article Title: Peroxiredoxin 3 regulates breast cancer progression via ERK-mediated MMP-1 expression

doi: 10.1186/s12935-024-03248-x

Figure Lengend Snippet: PRDX3 regulates MMP1 via MAPK. ( A ) Multi-pathway reporter array showed the transcriptional activities change of AP-1 in PRDX3-OE group (refer to Supplementary Figure for full investigated pathways). Relative luciferase activity was denoted as the ratio of firefly luciferase to renilla luciferase values. ( B ) Representative western blots showing protein expression of p-c-Jun, total c-Jun. Lamin B1 was used as nuclear loading control while GAPDH was used as cytoplasmic fraction control. ( C ) Human phospho-MAPK proteome profiler array demonstrates PRDX3 induced phosphorylation of MAPK and other serine/threonine kinases. Data is representative of a single experiment out of two experiments done. ( D ) Graph bars depicted the relative protein expression levels of several phosphorylated proteins in PRDX3-OE samples. ( E ) Treatment with SCH772984, an ERK inhibitor, for 24 h inhibited MMP1 expression in PRDX3-overexpressing MDA-MB-231 cells. Each bar represents mean ± SEM, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Relative expression levels of phosphorylated MAPK and other serine/threonine kinases were examined using the Proteome Profiler human phospoho-MAPK antibody array kit (R&D Systems).

Techniques: Luciferase, Activity Assay, Western Blot, Expressing, Control, Phospho-proteomics

Journal: Cells

Article Title: LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk

doi: 10.3390/cells9112522

Figure Lengend Snippet: LIMD2 knockout significantly reduced phosphorylation of nearly 60 kinases. The Human Phospho-MAPK and Human Phospho-Kinase Antibody Array Kit were assessed in LIMD2-edited cells (BCPAP E and TPC1 E ) and parental cells. ( A ) BCPAP P ; ( B ) TPC1 P . Log graph based on the fold change of all kinases analyzed in the proteome profiler array, showing that the LIMD2 KO reduced the phosphorylation levels of the majority of kinases analyzed.

Article Snippet: For the parallel determination of the relative phosphorylation of 65 multiple kinases associated with the three major families of MAPK, cells lysates were incubated with proteome profiler Human Phospho-MAPK array kit (ARY002B) and the proteome profiler Human Phospho-Kinase Array Kit (ARY003B) (R&D Systems, Minneapolis, MN, Canada), as previously described [ ].

Techniques: Knock-Out, Ab Array