Anti phospho thr202 tyr204 erk 1 2

Manufactured by Cell Signaling Technology

Sourced in United Kingdom

Anti-phospho-Thr202/Tyr204-Erk 1/2 is a lab equipment product that specifically detects phosphorylation of Thr202 and Tyr204 in the activation loop of Erk1/2 (p44/42 MAPK).

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Lab products found in correlation

Close spelling variants of this product

Phospho-ERK (527 citations) Anti-ERK (519 citations) Anti-phospho-ERK (265 citations) ERK/phospho-ERK (3 citations)

8 protocols using anti phospho thr202 tyr204 erk 1 2

Antibody Validation for Cell Signaling

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Anti-Sema6A (ab72369) was from Abcam (Cambridge, UK), anti-Lamin A (#86846), anti-Akt (#9272), anti-phospho-ser473-Akt (#9271), anti-Erk1/2 (#9102), anti-phospho-Thr202/Tyr204-Erk 1/2 (#9101), anti-YAP (#12395), anti-phospho-ser127-YAP (#13008), anti-p65 (#8242), anti-phospho-ser536-p65 (#3033), anti-Tubulin (#2125) were from Cell Signaling (Danvers, MA USA), anti-Hsp-70 (ab-83,392) and anti-GAPDH (ab-81,594) were from Immunological Sciences (Rome, Italy). HRP-conjugated secondary antibodies were from Bio-Rad (Hercules, CA USA).

Loria R., Laquintana V., Scalera S., Fraioli R., Caprara V., Falcone I., Bazzichetto C., Di Martile M., Rosanò L., Del Bufalo D., Bossi G., Sperduti I., Terrenato I., Visca P., Soddu S., Milella M., Ciliberto G., Falcioni R., Ferraresi V, & Bon G. (2022). SEMA6A/RhoA/YAP axis mediates tumor-stroma interactions and prevents response to dual BRAF/MEK inhibition in BRAF-mutant melanoma. Journal of Experimental & Clinical Cancer Research : CR, 41, 148.

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Western Blot Analysis of Kinase Activation

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Protein from co-cultured cells was extracted using the protease inhibitor cocktail and phosphatase inhibitor cocktail (Kangchen Bio-tech, China). Cell lysates were subjected to SDS-PAGE electrophoresis and proteins were blotted onto PVDF membranes. Appropriate primary antibodies and corresponding HRP-conjugated secondary antibodies were used. Exposure was performed using enhanced chemiluminescence reagents (Millipore, Billerica, MA, USA) and x-ray film. Antibodies used in western blotting were: anti-p38, anti-phospho-Thr180/Tyr182 p38, anti-ERK1/2, anti-phospho-Thr202/Tyr204 ERK1/2, anti-JNK, anti-phospho-Thr183/Tyr185 SAPK/JNK, and anti-GAPDH (all from Cell Signaling Technology).

Wu M., Chen X., Lou J., Zhang S., Zhang X., Huang L., Sun R., Huang P., Wang F, & Pan S. (2016). TGF-β1 contributes to CD8+ Treg induction through p38 MAPK signaling in ovarian cancer microenvironment. Oncotarget, 7(28), 44534-44544.

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Drosophila Protein Expression Analysis

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Drosophila were harvested and snap-frozen after aging and drug treatment, and tissue was homogenized and normalized for protein content using lysate (Ratliff et al., 2016 (link)). Antibodies used were as follows: Anti-phospho-Thr398-S6K (Cell Signaling #9209, United States); anti-S6K (Cell Signaling #9202, United States); anti-phospho-Thr308-Akt (Thermo Fisher Scientific 44-602G, United States); anti-Akt (Cell Signaling #9272, United States); anti-phospho-Thr37/46-4E-BP1 (Cell Signaling #2855, United States);anti-4E-BP1 (Cell Signaling #9452, United States); anti-ERK1/2 (Cell Signaling #4695, United States); anti-phospho-Thr202/Tyr204-ERK1/2 (Cell Signaling #9101, United States); anti-beta Tublin (abcam ab179513, Britain); horseradish peroxidase conjugated anti-mouse IgG (Cell Signaling #7076, United States) or horseradish peroxidase-conjugated anti-rabbit IgG (ProteinTech SA00001-2, United States). Nitrocellulose filter membranes with attached protein bands were imaged using the iBright FL1000 Imaging System (Invitrogen; Thermo Fisher Scientific, Inc., United States). Image J software 1.53a (National Institutes of Health, United States) was used to quantify the gray value of western blot bands and protein expression level was normalized as the gray value of the target protein/the loading control protein.

Zhao Q., Liu Y., Zhang S., Zhao Y., Wang C., Li K., Jin Z., Qiao J, & Liu M. (2022). Studies on the Regulation and Molecular Mechanism of Panax Ginseng Saponins on Senescence and Related Behaviors of Drosophila melanogaster. Frontiers in Aging Neuroscience, 14, 870326.

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Dopaminergic and Cholinergic Neurotransmission

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SKF 81297 and quinpirole were from Tocris (Minneapolis, MN, USA), dicyclomine, 6-OHDA-HCl, L-DOPA (3-(3,4-Dihydroxyphenyl-2,5,6-d₃)-L-alanine), desipramine, pargyline and benserazide were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-tyrosine hydroxylase (RRID: AB_390204) and anti-Choline Acetyltransferase (RRID:AB_2079751) and anti-pS10H3 (RRID:AB_1977177) were purchased from Millipore. Anti-phospho-Thr202/Tyr204–Erk1/2 (RRID:AB_331646), and phospho-(Ser/Thr) PKA substrate (RRID:AB_331817), a-DARPP-32 (RRID:AB_823479).
And anti pS6 ribosomal protein (RRID:AB_916156) were from Cell Signaling Technology. Various Alexa Fluor antibodies were used at a 1:500 dilution for immunohistochemistry (Invitrogen).

Castello J., Cortés M., Malave L., Kottmann A., Sibley D.R., Friedman E, & Rebholz H. (2020). The Dopamine D5 receptor contributes to activation of cholinergic interneurons during L-DOPA induced dyskinesia. Scientific Reports, 10, 2542.

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Capsaicin-Induced Pain in CDKL5 Mice

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Capsaicin (10 μl) [0.1 g/ml⁻¹ 100% ethanol; 1:1000 in PBS, 0.5% Tween 80, 10% ethanol] or vehicle (PBS, 0.5% Tween 80, 10% ethanol) was administered via intraplantar injection (69 ) to adult WT and Cdkl5 mutant mice or to adult Cdkl5-Floxed mice treated, two months before, with AAV5-GFP or AAV5-GFP-CRE (intra-sciatic injection). After capsaicin injection, the licking time was measured during the first 3 minutes. Finally, WT vs Cdkl5 null adult mice were sacrificed to study the activation of the peripheral pain pathways in fixed sciatic DRG and spinal cord. Immunohistochemistry on DRG and spinal cord sections was performed by using anti-phospho-Thr286-CaMKIIα (mouse, Thermo-Fisher MA1-047, 1:100), or anti-phospho-Thr202/Tyr204-Erk1/2 (rabbit, Cell Signaling #9101, 1:100).

La Montanara P., Hervera A., Baltussen L., Hutson T.H., Palmisano I., De Virgiliis F., Kong G., Chadwick J., Gao Y., Bartus K., Majid Q.A., Gorgoraptis N., Wong K., Downs J., Pizzorusso T., Ultanir S., Leonard H., Yu H., Millar D.S., Istvan N., Mazarakis N.D, & Giovanni S.D. (2020). Cyclin-dependent-like kinase 5 is required for pain signaling in human sensory neurons and mouse models. Science translational medicine, 12(551), eaax4846.

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Protein Analysis of HUVEC Lysates

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Total cell lysates of HUVECs were prepared as described previously [39 (link)]. Crude membrane fractions were prepared as previously [40 (link)], and solubilized on ice for 30 min with 1% NP-40. After mixing with Laemmli buffer, SDS-polyacrylamide gel electrophoresis and western blot analysis were performed [39 (link)]. The antibody-treated PVDF membrane was developed with ECL Prime Western Blotting Detection System and imaged by Amersham Imager 680 (GE Healthcare).
Primary antibodies used are as follows: anti-Na⁺/K⁺-ATPase α1 (sc-21712), anti-p70S6K (sc-230), anti-Akt (sc-1618), anti-Src (sc-8056), and anti-PLCγ (sc-7290) from Santa Cruz Biotechnology; anti-phospho-Thr389-p70S6K (9243), anti-S6 ribosomal protein (2217), anti-phospho-Ser235/Ser236-S6 ribosomal protein (4858), anti-eIF2α (eukaryotic initiation factor 2α subunit) (5324), anti-phospho-Ser51-eIF2α (3398), anti-phospho-Thr308-Akt (13038), anti-phospho-Ser473-Akt (4060), anti-Erk1/2 (4696), anti-phospho-Thr202/Tyr204-Erk1/2 (4370), anti-VEGFR2 (9698), anti-phospho-Tyr1175-VEGFR2 (2478), anti-p38 (8690), anti-phospho-Thr180/Tyr182-p38 (4511), anti-phospho-Tyr416-Src (6943), anti-FAK (3285), anti-phospho-Tyr397-FAK (8556), and anti-phospho-Ser1248-PLCγ (8713) from Cell Signaling Technology; anti-LAT1 (KE026) from TransGenic; anti-β-actin (66009–1-Ig) from Proteintech.

Quan L., Ohgaki R., Hara S., Okuda S., Wei L., Okanishi H., Nagamori S., Endou H, & Kanai Y. (2020). Amino acid transporter LAT1 in tumor-associated vascular endothelium promotes angiogenesis by regulating cell proliferation and VEGF-A-dependent mTORC1 activation. Journal of Experimental & Clinical Cancer Research : CR, 39, 266.

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Immunoblotting for Protein Expression

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For protein expression analysis, a standard immunoblotting protocol was followed as described previously (10 (link)). The antibodies used in this study were: anti-GDF15 (Santa Cruz Biotechnology), anti-RhoB total (Abcam), anti-phospho (Thr202/Tyr204) Erk1/2 (Cell Signaling Technology), anti-total Erk1/2 (Cell Signaling Technology), anti-phospho-c-Jun (S63, Abcam), and anti-total-c-Jun (Abcam). Antibodies against β-actin or β-tubulin were used as loading control.

Kalli M., Voutouri C., Minia A., Pliaka V., Fotis C., Alexopoulos L.G, & Stylianopoulos T. (2019). Mechanical Compression Regulates Brain Cancer Cell Migration Through MEK1/Erk1 Pathway Activation and GDF15 Expression. Frontiers in Oncology, 9, 992.

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Antibody sources for cellular analysis

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Anti-Sema6A (ab72369) was from Abcam (Cambridge, UK), anti-Lamin A (#86846), anti-Akt (#9272), anti-phospho-ser473-Akt (#9271), anti-Erk1/2 (#9102), anti-phospho-Thr202/Tyr204-Erk 1/2 (#9101), anti-YAP (#12395), anti-phospho-ser127-YAP (#13008), anti-p65 (#8242), anti-phospho-ser536-p65 (#3033), anti-Tubulin (#2125) were from Cell Signaling (Danvers, MA USA), anti-Hsp-70 (ab-83392) and anti-GAPDH (ab-81594) were from Immunological Sciences (Rome, Italy). HRP-conjugated secondary antibodies were from Bio-Rad (Hercules, CA USA).

Loria R., Laquintana V., Scalera S., Fraioli R., Caprara V., Falcone I., Bazzichetto C., Martile M.D., Rosanò L., Bufalo D.D., Bossi G., Sperduti I., Terrenato I., Visca P., Soddu S., Milella M., Ciliberto G., Falcioni R., Ferraresi V., & Bon G. (2021). SEMA6A/RhoA/YAP Axis Mediates Tumor-Stroma Interactions and Prevents Response to Dual BRAF/MEK Inhibition in BRAF-Mutant Melanoma.

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