P44 42 mapk erk1 2 l34f12

Manufactured by Cell Signaling Technology

Sourced in United States

P44/42 MAPK (Erk1/2) (L34F12) is a primary antibody that recognizes the extracellular signal-regulated kinases (ERK1/2), also known as p44/42 mitogen-activated protein kinases (MAPK). This antibody is used for the detection and analysis of ERK1/2 proteins in various research applications.

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Lab products found in correlation

Bca protein assay, by Thermo Fisher Scientific (4 mentions) Ripa buffer, by Thermo Fisher Scientific (4 mentions) Enhanced chemiluminescence, by Bio-Rad (2 mentions) Phospho p44 42 mapk t202 y204, by Cell Signaling Technology (2 mentions) Parp and cleaved parp, by Cell Signaling Technology (2 mentions) Chemidoc xrs system, by Bio-Rad (2 mentions) Image lab software, by Bio-Rad (2 mentions) P akt ser473 d9e xp, by Cell Signaling Technology (1 mentions) Gfp tag, by Proteintech (1 mentions) Gw4869, by Merck Group (1 mentions) β actin, by Proteintech (1 mentions) Odyssey sa infrared imaging system, by LI COR (1 mentions) Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions) Image studio, by LI COR (1 mentions)

Close spelling variants of this product

Mouse anti-p44/42 MAPK (Erk1/2) (L34F12) Anti- P44/42 MAPK (ERK1/2) (L34F12) Phospho-p44/42 MAPK (ERK1/2) (Thr 202/Tyr 204) (L34F12) P44/42 MAPK (Erk1/2) ERK1/2 p44/42 P44/42 MAPK ERK1/2 MAPK ERK1/2

4 protocols using p44 42 mapk erk1 2 l34f12

Protein Analysis via Western Blotting

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For protein analysis, cells were resuspended in a lysis RIPA buffer (#89901, Thermo, IL, USA) containing Phosphatase (#78426, Thermo, IL, USA) and Protease (#78429, Thermo) inhibitors cocktails at 4 °C, followed by the BCA protein assay (#23225, Thermo, IL, USA). Protein samples were separated electrophoretically by 15% SDS–PAGE and were then transferred to a polyvinylidene difluoride membrane (#ISEQ0001, Sigma-Aldrich, UK). The membranes were incubated with primary and secondary antibodies and then developed with enhanced chemiluminescence (#170-5061, Bio-Rad, CA, USA). The primary antibodies and their dilution factors were the 1 : 1000. For the western analysis, phospho-p44/42 MAPK (T202/Y204) (#4370S, Cell Signaling, MA, USA), p44/42 MAPK (Erk1/2) (L34F12) (#4696S, Cell Signaling, MA, USA), PARP and cleaved PARP (#9542S, Cell Signaling, MA, USA), tubulin (3873S, Cell Signaling, MA, USA) antibodies were used. For the detection of the signal from the membranes was used The ChemiDoc XRS+ System (Bio-Rad, USA) and for quantitative analysis of western blots was used Image Lab software (Bio-Rad, USA).

Gusakov E.A., Topchu I.A., Mazitova A.M., Dorogan I.V., Bulatov E.R., Serebriiskii I.G., Abramova Z.I., Tupaeva I.O., Demidov O.P., Toan D.N., Lam T.D., Bang D.N., Boumber Y.A., Sayapin Y.A, & Minkin V.I. (2021). Design, synthesis and biological evaluation of 2-quinolyl-1,3-tropolone derivatives as new anti-cancer agents. RSC Advances, 11(8), 4555-4571.

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Measuring Ras Signaling Pathway

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The following antibodies used to measure Ras signaling were purchased from Cell Signaling Technology (Danvers, MA): pAkt (Ser473) (D9E) XP (Cat#4060), total Akt (pan) (40D4) (Cat#2920), p-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP (Cat#4370) and p44/42 MAPK (Erk1/2) (L34F12) (Cat#4696). The following antibodies used to measure housekeeping genes were purchased from Proteintech Group (Rosemont, IL): β-actin (Cat#60008-1-1 g) and GFP-tag (Cat#66002-1-lg; clone #1E10H7). cDNA of human SMPD1 (GenScript, Cat# OHu18710D), SMPD2 (GenScript, Cat# OHu30447D) and SMPD3 (Sino Biological, Cat# HG15755-G) were cloned into GFP-N1 vector. CellLight Lysosomes-RFP (Cat#C10597), Early endosomes-RFP (Cat#C10587), Late Endosomes-RFP (Cat#C10589), Golgi-RFP (Cat#10593), Mitochondria-RFP (Cat#C10601) and ER-tracker (E34250) were purchased from Invitrogen. Altenusin (Cat#SML2193) and GW4869 (Cat#567715) were purchased from MilliporeSigma. Avicin compounds were provided in collaboration with Dr. Jordan Gutterman (MD Anderson Cancer Center; Houston, TX).

Garrido C.M., Henkels K.M., Rehl K.M., Liang H., Zhou Y., Gutterman J.U, & Cho K.J. (2020). Avicin G is a potent sphingomyelinase inhibitor and blocks oncogenic K- and H-Ras signaling. Scientific Reports, 10, 9120.

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MAPK1/3 and RAS Protein Detection

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Cells were lysed with 500 µL RIPA-Lysis buffer supplemented with phosphatase and proteinase inhibitors [53 (link)], and homogenized utilizing a 0.45 × 25 mm needle on a syringe. Debris was removed by centrifugation at 16,100× g for 15 min at 4 °C. Subsequently, 50 µg of protein was applied for Western blotting. For total-MAPK1/3 detection, p44/42 MAPK (Erk1/2) (L34F12) (#4696), and for phospho-MAPK1/3, rabbit anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP (#4370), from Cell Signaling were used. RAS was detected using pan-RAS (Ab-3) (#OP40) from Merck (Darmstadt, Germany). Protein detection was performed using the Odyssey Sa Infrared Imaging System (LI-COR Bioscience, Lincoln, NE, USA) or enhanced chemiluminescence (ECL, Thermo Scientific, Waltham, MA, USA). ImageStudio (LI-COR) software was employed for quantitative analysis.

Stieglitz F., Gerhard R., Hönig R., Giehl K, & Pich A. (2022). TcdB of Clostridioides difficile Mediates RAS-Dependent Necrosis in Epithelial Cells. International Journal of Molecular Sciences, 23(8), 4258.

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Protein Analysis by Western Blotting

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For protein analysis, cells were resuspended in a lysis RIPA buffer (#89901, Thermo, IL, USA) containing Phosphatase (#78426, Thermo, IL, USA) and Protease (#78429, Thermo) inhibitors cocktails at 4 °C, followed by the BCA protein assay (#23225, Thermo, IL, USA). Protein samples were separated electrophoretically by 15% SDS–PAGE and were then transferred to a polyvinylidene difluoride membrane (#ISEQ0001, Sigma-Aldrich, UK). The membranes were incubated with primary and secondary antibodies and then developed with enhanced chemiluminescence (#170–5061, Bio-Rad, CA, USA). The primary antibodies and their dilution factors were the 1 : 1000. For the western analysis, phospho-p44/42 MAPK (T202/Y204) (#4370S, Cell Signaling, MA, USA), p44/42 MAPK (Erk1/2) (L34F12) (#4696S, Cell Signaling, MA, USA), PARP and cleaved PARP (#9542S, Cell Signaling, MA, USA), tubulin (3873S, Cell Signaling, MA, USA) antibodies were used. For the detection of the signal from the membranes was used The ChemiDoc XRS+ System (Bio-Rad, USA) and for quantitative analysis of western blots was used Image Lab software (Bio-Rad, USA).

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