ERK1 + ERK2

Protein Overview

ERK1/2 (MAPK3/MAPK1) are the terminal effector kinases of the MAPK/ERK cascade, phosphorylating over 150 identified substrates involved in transcription, translation, and cytoskeletal dynamics. This total (pan) antibody detects ERK1/2 regardless of phosphorylation state, serving as an expression/loading control for phospho-ERK studies.

Expected Localization

Present constitutively in the cytoplasm, with additional pools at the plasma membrane and low steady-state nuclear presence.

Localization shifts toward the nucleus upon activation, but total protein is detectable in both compartments regardless of stimulation.

Expected Expression Pattern

Ubiquitously and constitutively expressed across virtually all mammalian cell and tissue types, with stable total protein levels that do not require stimulation to detect and do not change acutely with growth factor treatment.

Relevant Pathways

RAS/RAF/MEK/ERK (canonical MAPK) cascade
Receptor tyrosine kinase signaling (EGFR, FGFR, PDGFR)
Cell proliferation, differentiation, and survival
Transcriptional regulation (Elk-1, c-Fos, c-Myc substrates)
Cytoskeletal reorganization and cell migration

HeLa cells were cultured in a 12-well format Cytos slide kit overnight, fixed with 4% formaldehyde for 30 minutes, permeabilized using 70% ethanol for 10 minutes, and then incubated with probe made with biotinylated rabbit monoclonal anti-ERK1 + ERK2 antibody for 1 hour at room temperature. Negative control probe based on Biotin Rabbit Isotype Control (ABclonal AC042BC) was included to assess background signal and potential crosstalk, with no detectable crosstalk observed. The slide kit was then assembled into a flow cell and loaded onto an AVITI24™ along with the Cytos Protein Cartridge for a multiomics cell screening run. ERK1 + ERK2 demonstrated strong cytoplasmic localization and high signal-to-noise ratio, with no detectable probe crosstalk. These results support high confidence in target-specific detection and validate this antibody as a passing clone. A549 cells were also cultured and processed under the same protocol in parallel as part of Cytos cross-run reproducibility testing.

Signal-to-noise ratio
Acceptance criteria: > 2x background

Pass

10.6 ± 1.5 (HeLa; n=22 runs)
10.0 ± 2.3 (A549; n=36 runs)

Indicates the fold brightness of the target signal as measured by the Mean Intensity Above (Otsu-defined) Threshold (IAT), over the background (a channel-matched isotype control) in the same cell. Data shown are mean ± standard deviation across independent reproducibility runs. The signal is consistently around 5 times above the acceptance threshold in both HeLa and A549 cells — a clean, easily distinguishable signal.

Panel-level run-to-run reproducibility
Acceptance criteria: R² > 0.9

Pass

For each imaging panel, we assess reproducibility by comparing the intensity signal a given antibody target produces across independent replicate runs. Within each run, we measure the average per-well fluorescence intensity for every target in the panel. We then plot each target's intensity from one run against the matching target's intensity from another run of the same layout, and calculate the Pearson correlation coefficient (r) between those two sets of measurements. Squaring that value gives the R² (coefficient of determination), which reports what fraction of the variation in one run's signal is explained by the other run's signal.

Protein Overview

Clone Information

Qualitative Results

Quantitative Results

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