To inhibit the EGFR signaling pathway, 70% confluent BT-549 and MDA-MB-231 cells were cultured with serum free medium overnight followed by a 5-hour incubation in medium containing 10 μM EGFR small molecule inhibitor Erlotinib (Cell Signaling Technology, Beverly, MA). Epidermal growth factor receptor inhibition was verified by immunoblotting with rabbit anti-Phospho-EGF Receptor (Tyr1068) (Cell Signaling Technology). The PI3K pathway was analyzed using rabbit anti-p-Akt and rabbit anti-Akt antibodies (Cell Signaling Technology), and mitogen-activated protein kinase (MAPK) pathway inhibition by immunoblotting with a rabbit anti-p-ERK 1/2 antibody (Cell Signaling Technology) and a rabbit anti-ERK 2 antibody (Santa Cruz Biotechnology, Santa Cruz, CA).
Rabbit anti erk2 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Rabbit anti-ERK2 antibody is a primary antibody that specifically recognizes the extracellular signal-regulated kinase 2 (ERK2) protein. It is used for the detection and quantification of ERK2 in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Erlotinib, by Cell Signaling Technology (1 mentions)
Rabbit anti perk1 2 antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti phospho egf receptor tyr1068, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Daam1, by Proteintech (1 mentions)
Rabbit anti erk1, by Santa Cruz Biotechnology (1 mentions)
Fhod1, by Merck Group (1 mentions)
Rabbit anti p44 42 mapk erk1 2 antibody, by Cell Signaling Technology (1 mentions)
3 protocols using rabbit anti erk2 antibody
1
EGFR Signaling Inhibition in Breast Cancer Cells
2
Flag-Protein Expression Analysis in HEK293T
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Transfected HEK293T cells were collected in phosphate buffered saline (PBS), pelleted, and lysed in solubilization buffer (50 mM Tris–HCl pH 8, 1% Igepal and 1 mM EDTA) with protease inhibitor cocktail (Sigma Aldrich). The cell lysate was centrifuged at 14,000×g for 20 min. Equal amounts of protein were separated in 10% SDS-PAGE. The resolved proteins were electroblotted to PVDF membranes, and the blots were blocked with 5% non-fat dry milk in PBS-T (Tween 0.1%) and incubated overnight at 4°C with the anti-Flag antibody conjugated to peroxidase (1:10000). Immunoreactive bands were detected by using ECL Plus Western Blotting Substrate. ERK2, detected by a rabbit anti-ERK2 antibody (Santa Cruz, USA), was used as loading control. Densitometric analysis was achieved with ImageJ software.
3
Verifying siRNA and Pathway Inhibition
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Western blot samples were collected and processed as previously described [9 (link)]. To verify the efficacy of the siRNA treatments, we used rabbit anti-HER2/ERBB2 (Proteintech), FHOD1 (Sigma-Aldrich), INF2, and DAAM1 (both Proteintech) antibodies. PI3K/Akt pathway inhibition was verified by immunoblotting using rabbit anti-pAkt and Akt antibodies (Cell Signaling Technology), and MEK/ERK pathway inhibition was verified by immunoblotting with a rabbit anti-p44/42 MAPK (Erk1/2) antibody (Cell Signaling Technology) and rabbit anti-ERK1 mixed with rabbit anti-ERK2 antibody (both from Santa Cruz Biotechnology, USA). All primary antibodies were used at 1:1000 dilution. The secondary antibodies used were horseradish peroxidase-conjugated swine anti-rabbit or rabbit anti-mouse (Agilent, USA) diluted 1:3,000 in blocking solution. The control for protein loading was CoraLite® Plus 488-conjugated glyceraldehyde 3-phosphate dehydrogenase mouse monoclonal antibody (Proteintech) at 1:5,000.
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