α p44 42 mapk erk1 2

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The α-p44/42 MAPK (ERK1/2) is a primary antibody that recognizes the phosphorylated forms of the extracellular signal-regulated kinase 1 and 2 (ERK1/2) proteins. ERK1/2 are mitogen-activated protein kinases (MAPK) that play a central role in the MAPK/ERK signaling pathway.

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ERK1/2 (1487 citations) P44/42 MAPK (Erk1/2) (135 citations) P44/42 MAPK (90 citations) α-ERK1/2 (10 citations) ERK1/2 MAPK (3 citations) ERK1/2 p44/42 (2 citations)

4 protocols using α p44 42 mapk erk1 2

Immunostaining and Protein Analysis of RhoGTPases

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The following antibodies were used for immunostaining: α-VE-cadherin XP (#2500; Cell Signaling Technologies, Danvers, MA) and a-RhoB (#sc-8048; Santa Cruz Biotechnology, Santa Cruz, CA). Alexa Fluor 488 secondary antibody (anti-rabbit and anti-mouse) and Alexa Fluor 647 secondary antibody (anti-rabbit) (all Invitrogen) were the secondary antibodies.
The following antibodies were used for protein analysis: ß-actin (Merck), LDLR (Biovision), and SQLE (Proteintech). α-p44/42 MAPK (ERK1/2) (#9102), α-RhoA (#2117), α-RhoC (#3430), and α-GAPDH (#2118; Cell Signaling Technologies), α-RhoB (#sc-180, #sc-8048; Santa Cruz Biotechnology), α-HA (H3663), and α-vinculin (V4139; Sigma-Aldrich), α-Fbxw7 (ab171961; Abcam) and mouse α-ubiquitin (FK-2; Boston Biochem). HRP-conjugated goat–anti-rabbit antibody and goat–anti-mouse (Dako) were used as secondary antibodies.
For inhibition of geranylgeranylation and farnesylation, GGTI-298 and FTI-277 HCL (both Selleck Chemicals) were used. For normalization of prenylation in the FBXW7 knockdown cells, GGPP or FPP were applied (Sigma).
The siRNAs used were ON-TARGET plus Nontargeting pool (siNT), ON-TARGET plus Human FBXW7 pool (siFBXW7), ON-TARGET plus Human RHOA siRNA pool (siRhoA), ON-TARGET plus Human RHOB siRNA pool (siRhoB), and ON-TARGET plus Human FBXW7 siRNA—Set of 4 Upgrade (all Dharmacon/GE-Healthcare, Lafayette, CO).

Pronk M.C., Majolée J., Loregger A., van Bezu J.S., Zelcer N., Hordijk P.L, & Kovačević I. (2019). FBXW7 regulates endothelial barrier function by suppression of the cholesterol synthesis pathway and prenylation of RhoB. Molecular Biology of the Cell, 30(5), 607-621.

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Protein Expression Profiling by Western Blot

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Cells were lysed using lysis buffer (40 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% sodium dodecyl sulfate (SDS)). Approximately 20 μg of the protein extract was resolved via SDS-PAGE and analyzed via immunoblotting with primary antibodies against α-HIF-1α (Cell Signaling, Danvers, MA, USA, 3716), α-PARP (Cell Signaling, 9542), α-Caspase-3 (Cell Signaling, 9662), α-p44/42 MAPK (ERK1/2) (Cell Signaling, 4695), α-Phospho-p44/42 MAPK (Erk1/2) (Cell Signaling, 4370), α-p38 MAPK (Cell Signaling, 9212), α-Phospho-p38 MAPK (Cell Signaling, 4511), α-JNK2 (Cell Signaling, 9258), α-Phospho-SAPK/JNK (Cell Signaling, 4668), α-Cyclin B1 (Santa Cruz, Dallas, TX, USA, sc-752), α-Cyclin D1 (Cell Signaling, 2978), α-Cyclin E1 (Cell Signaling, 4129), α-CDK4 (Cell Signaling, 12790), α-p53 (Millipore, Burlington, MA, USA, 05-224), α-p21 (Cell Signaling, 2947), α-TBP (Abcam, Cambridge, UK, ab818), α-β-actin (Santa Cruz, sc-47778), and α-alpha-tubulin (Abcam, ab7291).

Kim W.H., Kim M.J., Jin J.O, & Lee P.C. (2023). IDF-11774 Induces Cell Cycle Arrest and Apoptosis by Inhibiting HIF-1α in Gastric Cancer. Pharmaceutics, 15(12), 2772.

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Comprehensive Antibody Evaluation for Cell Signaling

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Antibodies against α-Rab5a (#2143), α-Rab7a (#9367), α-Rab10 (#8127), α-EGFR (#4267), α-Phospho-EGF Receptor (Tyr1068) (#2234 S), α-Phospho-p38 MAPK (Thr180/Tyr182) (#2775 S), α-p38 MAPK (D13E1) XP Rabbit mAb (#8690), α-p44/42 MAPK (Erk1/2) (#695), α-IRG1 (#17805) were purchased from Cell Signaling Technology. Antibodies against α-HIF1-α (sc-53546), α-EGFR (sc-120), α-PKM2 (sc-365684), α-LDHA (sc-133123), α-MAPK14 (sc-11415), α-EGFR (sc-120), α-EEA1 (sc-137130), α-TGN38 (sc-166594), α-LAMP1 (sc-20011), α-PPARγ (sc-7273), α-SDHA (sc-390381), α-COX2 (sc-514489), α-COX4 (sc-517553), α-ATP5A (sc-136178) were purchased from Santa Cruz Biotechnology. Antibodies against α-HA (51064-2-AP, proteintech Group), α-FLAG (20543-1-AP, proteintech Group), APC anti-mouse CD11b (APC-65055, proteintech Group) were purchased from proteintech Group. Antibodies against APC anti-mouse iNOS (#85-17-5920-82, eBioscience), PE anti-mouse CD206 (MMR) Antibody (#85-12-2061-82, eBioscience), α-F4/80 (11-4801-82, FITC, eBioscience), PE anti-mouse Ly6G (#12-9668-82, eBioscience), α-CD14 (25-0149-42, FITC, eBioscience) were purchased from eBioscience. Antibodies against α-GAPDH (T0004), α-β-Tubulin (T0023), α-Phospho-PPAR gamma (Ser112) (AF3284), α-Phospho-PKM2 (AF7231) were purchased from Affinity Biosciences.

Zhang X., Chen C., Ling C., Luo S., Xiong Z., Liu X., Liao C., Xie P., Liu Y., Zhang L., Chen Z., Liu Z, & Tang J. (2022). EGFR tyrosine kinase activity and Rab GTPases coordinate EGFR trafficking to regulate macrophage activation in sepsis. Cell Death & Disease, 13(11), 934.

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Investigating Cell-Cell Adhesion Regulators

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The following antibodies were used in this study: αVE-cadherin XP (#2500), αRhoA (#2117), αRhoC (#3430), αCullin-3 (#2759), αp44/42 MAPK (ERK1/2) (#9102), αpMLC2 Ser19 (#3671), αGAPDH (#2118), αIκBα (#4841) and αpp65 (#93H1) (all from Cell Signaling Technology); αRhoB (#sc-8048 and #sc-180), αCullin-1 (#sc-17775), αICAM (#sc-8439) (all from Santa Cruz Biotechnology); αCullin-2 (#610778) and αp38 (#61268) (BD Transduction Laboratories) and αpp38 (#09–272) (Millipore).
Horseradish peroxidase-conjugated goat anti-rabbit and anti-mouse antibodies (Dako) were used as secondary antibodies for western blotting. For immunofluorescent staining DAPI (Thermo Fisher scientific), Alexa 488-secondary antibody (anti-rabbit and anti-mouse, Invitrogen) and Acti-stain 670 phalloidin (Cytoskeleton) were used.
The following inhibitors and cytokines were used in this study: CSN5i-3 (Novartis), MLN4924 (Active Biochem), Y27632 (#Y0503) (Sigma), TNF-α (#300-01 A) (PeproTech), BAY11-7085 (Cayman Chemical).

Majolée J., Pronk M.C., Jim K.K., van Bezu J.S., van der Sar A.M., Hordijk P.L, & Kovačević I. (2019). CSN5 inhibition triggers inflammatory signaling and Rho/ROCK-dependent loss of endothelial integrity. Scientific Reports, 9, 8131.

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