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Phospho erk antibody

Manufactured by Cell Signaling Technology
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The Phospho-ERK antibody is a laboratory reagent used to detect and quantify the phosphorylated form of the extracellular signal-regulated kinase (ERK) protein. ERK is a key signaling molecule involved in various cellular processes, and its phosphorylation is an important indicator of cellular activation and signaling pathway activation.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Total erk antibody, by Cell Signaling Technology (3 mentions) Phospho p38 antibody, by Cell Signaling Technology (3 mentions) Erk antibody, by Cell Signaling Technology (3 mentions) U0126, by Cell Signaling Technology (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Phospho jnk antibody, by Cell Signaling Technology (2 mentions) Antigen unmasking solution, by Vector Laboratories (2 mentions) Cd11b, by BD (2 mentions) F4 80 bm8, by Thermo Fisher Scientific (2 mentions) E cadherin, by BD (2 mentions) Pi3k antibody, by Santa Cruz Biotechnology (2 mentions) P38 antibody, by Cell Signaling Technology (2 mentions) Psb0739, by Bio-Techne (1 mentions) Total erk, by Cell Signaling Technology (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Adenosine diphosphate (adp), by Merck Group (1 mentions) Mrs2179, by Bio-Techne (1 mentions) Rabbit anti human bcl 2, by Cell Signaling Technology (1 mentions) Mesadp, by Bio-Techne (1 mentions)

Close spelling variants of this product

Phospho-ERK (527 citations) Anti-phospho-ERK antibody (25 citations) ERK antibody (19 citations) Anti-p42/44-phospho-ERK antibody (4 citations) ERK/phospho-ERK (3 citations) Rabbit anti-phospho-ERK polyclonal antibody (3 citations) Anti-phospho-ERK (Thr202/Tyr204) antibody (2 citations) Anti-phospho-ERK (Th202/Tyr204) antibody (2 citations) Primary antibody to phospho-ERK (2 citations) Anti‐phospho‐ERK primary antibody (2 citations)

20 protocols using phospho erk antibody

1

Nucleolin Regulation by Purinergic Signaling

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ADP, ATP, UDP, and UTP were purchased from Sigma-Aldrich (St. Louis, MO). Rabbit anti-human Bcl-2, total ERK, phospho-ERK antibodies, Rabbit anti-human nucleolin antibody, and ERK inhibitor U0126 were purchased from Cell Signaling Technology (Beverly, MA). P2Y1, 12, 13 agonist 2-MeSADP, P2Y1 selective inhibitor MRS2179, P2Y12 potential inhibitor PSB0739, P2Y13 competitive inhibitor MRS2211 were purchased from Tocris (Bristol, UK). Mammalian expression plasmid pReceive-M29 coding for eGFP-nucleolin fusion protein was purchased from GeneCopoeia (Germantown, MD).
Wang W., Luo J., Xiang F., Liu X., Jiang M., Liao L, & Hu J. (2014). Nucleolin Down-Regulation Is Involved in ADP-Induced Cell Cycle Arrest in S Phase and Cell Apoptosis in Vascular Endothelial Cells. PLoS ONE, 9(10), e110101.
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2

Apigenin Enhances NK Cell Activity

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Apigenin was purchased from Sigma-Aldrich. Recombinant human IL-2 (rhIL-2) and recombinant human IL-15 (rhIL-15) were purchased from Xiamen Amoytop Biotech Co., Ltd Stem cell growth medium (SCGM) was purchased from CellGenix GmbH. Dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from Thermo scientific. FITC-anti-human CD56, PerCP-Cy5.5-anti-human CD3, APC-anti-human CD107a, PE-anti-human Gran B, PE-anti-human Perforin, PE-anti-human NKG2D, and isotype controls were obtained from BD Biosciences. Trizol and SYBR Green PCR Master Mix were purchased from Thermo Fisher Scientific. PrimeScriptTMRT reagent Kit was purchased from Takara Bio Inc. The antibodies of Bcl-2, Bax, and GAPDH were obtained from Proteintech Group, Inc. phospho-SAPK/JNK and phospho-ERK antibodies were got from Cell Signaling Technology, Inc. AP-labed Goat anti-Rabbit IgG (H + L), AP-labed Goat anti-Mouse IgG (H + L), and CCK-8 kit were got from Beyotime Institute of Biotechnology.
Feng Y.B., Chen L., Chen F.X., Yang Y., Chen G.H., Zhou Z.H, & Xu C.F. (2023). Immunopotentiation effects of apigenin on NK cell proliferation and killing pancreatic cancer cells. International Journal of Immunopathology and Pharmacology, 37, 03946320231161174.
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3

Antibodies and Reagents for Cell Signaling

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Phospho-DRP1 (Ser616) and phospho-ERK antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). DRP1, β-tubulin and ERK1/2 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-EPHB6 was obtained from Santa Cruz and Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin (BSA) was acquired from BioShop Canada Inc. (Burlington, ON, Canada). Dimethyl sulfoxide (DMSO), puromycin, doxorubicin and polybrene were purchased from Sigma-Aldrich. Resazurin was obtained from R&D Systems (Minneapolis, MN, USA). PD0325901 was acquired from Tocris (Bio-Techne, Minneapolis, MN, USA). Tween-20 was obtained from Fisher Scientific (Ottawa, ON, Canada).
El Zawily A.M., Toosi B.M., Freywald T., Indukuri V.V., Vizeacoumar F.J., Leary S.C, & Freywald A. (2016). The intrinsically kinase-inactive EPHB6 receptor predisposes cancer cells to DR5-induced apoptosis by promoting mitochondrial fragmentation. Oncotarget, 7(47), 77865-77877.
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4

Protein Expression and Signaling Assay

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Propidium iodide (PI), Tris-HCl, trypan blue, ethylenediaminetetraacetic acid (EDTA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), ribonuclease A, and dimethyl sulfoxide (DMSO) were obtained from Sigma Chemical (St. Louis, MO, USA). Anti-HDA6 antibody was purchased from Novus Biologicals, LLC (Centennial, CO, USA). Anti-serpine-1, EGFR, β-catenin, vimentin, Cyclin-dependent kinase 2 (CDK2), fibronectin, Poly (ADP-ribose) polymerase (PARP), and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p38, phospho-p38, c-Jun N-terminal kinases (JNK), phospho-JNK, extracellular regulated protein kinases (ERK), and phospho-ERK antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-snail antibody was purchased form GenScript (Piscataway, NJ, USA). Anti-phospho-snail and Cox-2 antibodies were purchased from Abcam (Cambridge, MA, USA). Anti-cyclin A2 and cyclin B1 were purchased from GeneTex, Inc. (Irvine, CA, USA).
Wei P.L., Lin J.C., Hung C.S., Makondi P.T., Batzorig U., Chang T.C., Huang C.Y, & Chang Y.J. (2022). Human α-defensin 6 (HD6) suppresses CRC proliferation and metastasis through abolished EGF/EGFR signaling pathway. International Journal of Medical Sciences, 19(1), 34-46.
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5

Immunohistochemical Evaluation of Phospho-ERK and LIN28A

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Phospho-ERK antibody (Cell Signaling Technology) was used as described previously [38 (link)]. Anti-LIN28A (Cell Signaling Technology) was used as described previously [16 (link)]. AT/RT tissue cores were scored by a neuropathologist (CGE) using H-scores (H) (0–200) which were obtained by multiplying the intensity of stain (0: no stain, 1: weak stain, 2: strong stain) by percentage (0–100) of neoplastic cells showing the staining intensity. Tumors were scored for phospho-ERK and LIN28A as being negative (score of 0), low (5 to 45), intermediate (50-95) or high (100 and above).
Weingart M.F., Roth J.J., Hutt-Cabezas M., Busse T.M., Kaur H., Price A., Maynard R., Rubens J., Taylor I., Mao X.G., Xu J., Kuwahara Y., Allen S.J., Erdreich-Epstein A., Weissman B.E., Orr B.A., Eberhart C.G., Biegel J.A, & Raabe E.H. (2014). Disrupting LIN28 in atypical teratoid rhabdoid tumors reveals the importance of the mitogen activated protein kinase pathway as a therapeutic target. Oncotarget, 6(5), 3165-3177.
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6

Signaling Pathway Activation Analysis

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Alpha-minimum essential medium (MEM) and fetal bovine serum (FBS) were purchased from Gibco (Carlsbad, CA, USA). Trypsin, phosphate-buffered saline (PBS), and distilled water were purchased from Biowest (Carlsbad, CA, USA). Lipofectamine 3000 reagent was purchased from Invitrogen (Carlsbad, CA, USA). The extracellular signal-regulated kinase (ERK) inhibitor U0126, phospho-ERK antibody, total ERK antibody and peroxidase-labeled secondary antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies for hypoxia-inducible factor (HIF)-1α were purchased from BD Biosciences (Oxford, UK). Antibodies for GAPDH were provided by the Gwangju Institute of Science and Technology, Korea (GIST). The intracellular calcium chelatorBAPTA-AM was purchased from Calbiochem (La Jolla, CA, USA).
Choi W., Kwon S.J., Jin H.J., Jeong S.Y., Choi S.J., Oh W., Yang Y.S., Jeon H.B, & Jeon E.S. (2017). Optimization of culture conditions for rapid clinical-scale expansion of human umbilical cord blood-derived mesenchymal stem cells. Clinical and Translational Medicine, 6, 38.
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7

Western Blot Analysis of Signaling Pathways

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BPH-1 cells were lysed using Pro-prep TM protein extraction solution (iNtRON). Lysates were mixed with SDS-PAGE sample buffer and boiled at 100°C for 5 min. Equal amounts of protein samples were run on 10% SDS-PAGE gel and transferred onto PVDF membranes, which were incubated with the following primary antibodies (all at 1:1,000) overnight at 4°C: phospho-p38 antibody, phospho-JNK antibody, phospho-ERK antibody, anti-TLR4 antibody, phospho-STAT3 antibody (all from Cell Signaling, Beverly, Massachusetts, USA); also phospho-JAK2 antibody, phospho-NF-κB antibody, β-actin antibody (all from Abcam). After 3 washes with Tween 20-Tris buffered saline (T-TBS, Biosesang, Seongnam, Korea), proteins were probed with anti-rabbit HRP-conjugated secondary antibody 1:10,000 (Abcam). Proteins were visualized by enhanced chemiluminescence (ECL) using a ChemiDoc MP detection system (Bio-Rad, Hercules, California, USA).
Kim S.S., Kim J.H., Han I.H., Ahn M.H, & Ryu J.S. (2016). Inflammatory Responses in a Benign Prostatic Hyperplasia Epithelial Cell Line (BPH-1) Infected with Trichomonas vaginalis. The Korean Journal of Parasitology, 54(2), 123-132.
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8

Immunofluorescence Analysis of Tissue Specimens

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Tissue specimens were fixed in 10% formalin, dehydrated, and then embedded in paraffin. Tissue samples were cut at 5 µm thicknesses and stained with hematoxylin and eosin. For immunofluorescence analysis, tissue sections were subjected to antigen retrieval by boiling the slides in Antigen Unmasking Solution (Vector Laboratories) for 10 minutes according to instructions. Sections were then blocked for 1 hour at 22°C with 5% BSA in PBS and incubated overnight at 4°C with the primary antibodies (rabbit polyclonal Ki67, phospho-Stat3Tyr705 and phospho-Erk antibody from Cell Signaling used at a dilution of 1/100, and rat polyclonal CD63, Muc2 from AbCam [Cambridge, MA], rabbit polyclonal CD63, Jam-a from Santa Cruz Biotechnology Inc [Santa Cruz, CA] used at a dilution of 1/50, CD11b and E-Cadherin were gained from BD Bioscience (San Jose, CA) and used at a dilution of 1/100. Primary antibodies were detected by Alexa Fluor 488, 594 or 647 conjugated goat anti–mouse, anti–rabbit IgG and anti-rat (1:600, Invitrogen), respectively. Tissues were counterstained with DAPI and images were captured on a Zeiss LSM 510 confocal microscope equipped with a digital image analysis system (Pixera). For immunofluorescence analysis of macrophages, OCT (Sakura Finetek)-embedded tissue cryosections (9µm-thick) were stained by F4/80 (BM8, ebioscience).
Deng Z., Mu J., Tseng M., Wattenberg B., Zhuang X., Egilmez N.K., Wang Q., Zhang L., Norris J., Guo H., Yan J., Haribabu B., Miller D, & Zhang H.G. (2015). Enterobacteria-secreted particles induce production of exosome-like S1P-containing particles by intestinal epithelium to drive Th17-mediated tumorigenesis. Nature communications, 6, 6956.
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9

Western Blot Analysis of Phospho-ERK

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Transfected cells were treated with 10 nM E2. One hour prior to harvest, cells received a second E2 treatment, and were lysed in RIPA buffer containing protease and phosphatase inhibitors. Cell lysates were then centrifuged at 10 000g for 20 min to remove debris. Protein concentrations were measured using a BCA protein assay kit (Pierce). Proteins separated by SDS–PAGE were transferred onto nitrocellulose membranes. After blocking with blocking buffer (Tris-buffered saline, 0.05% Tween 20, 5% dry skim milk, pH 7.4) for 1 h at room temperature, membranes were incubated for 24 h with phospho-ERK antibody at 1 : 1000 dilution (Cell Signaling Technology, Beverly, MA). After incubation with secondary antibodies conjugated with HRP, proteins were visualized using an ECL detection kit (Pierce) according to the manufacturer's instructions. The membranes were then stripped with a stripping buffer (Pierce) and re-probed with total ERK antibody (Cell Signaling no. 9102). To control for total protein loading, membranes were also probed with an anti-tubulin antibody (Abcam). We used the following catalogue numbers and vendors:
Lin C.Y., Kleinbrink E.L., Dachet F., Cai J., Ju D., Goldstone A., Wood E.J., Liu K., Jia H., Goustin A.S., Kosir M.A., Thepsuwan P, & Lipovich L. (2016). Primate-specific oestrogen-responsive long non-coding RNAs regulate proliferation and viability of human breast cancer cells. Open Biology, 6(12), 150262.
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10

Peptide Effects on Hair Follicle Cells

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Example 3

Human hair follicle dermal papilla cells were seeded at a density of 4×105 cells/well on 6-well plates, followed by incubation overnight. After changing into serum-free medium, the cells were treated with the peptides, followed by incubation for 15 and 30 minutes, and then the wells were harvested to prepare the cell lysate. Western blotting was performed using PI3K antibodies (Santa Cruz Biotechnology, USA) and phospho-ERK antibody (Cell Signaling Technology, USA) to compare protein expression patterns. The results are shown in FIGS. 3a and 3b.

As can be confirmed from FIGS. 3a and 3b, the expression of PI3K and the phosphorylation of ERK, which influence the growth of human hair follicle dermal papilla, were increased by the treatment with the peptide composed of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

US10344061B2. Peptide exhibiting hair growth promoting activity and/or melanin production promoting activity and use thereof (2019-07-09). CAREGEN CO., LTD. [KR]. Inventors: Yong Ji Chung [KR], Eun Mi Kim [KR], Eung Ji Lee [KR].
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