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Phospho jnk1 2 thr183 tyr185

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Phospho-JNK1/2 (Thr183/Tyr185) is a laboratory reagent that detects the phosphorylation of JNK1 and JNK2 proteins at specific threonine and tyrosine residues. It is used to monitor the activation of these kinases in cellular signaling pathways.

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Lab products found in correlation

Phospho p38 thr180 tyr182, by Cell Signaling Technology (4 mentions) Phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (4 mentions) Anti tslp antibody, by Novus Biologicals (2 mentions) Firefly luciferase assay system, by Promega (2 mentions) H e staining kit, by Merck Group (2 mentions) Tnf α, by ProSpec (2 mentions) Total erk1 2, by Cell Signaling Technology (2 mentions) Total mek1 2, by Cell Signaling Technology (2 mentions) β mhc, by Santa Cruz Biotechnology (2 mentions) Total p38, by Cell Signaling Technology (2 mentions) Phospho mek1 2 ser217 221, by Cell Signaling Technology (2 mentions) Phosstop phosphatase inhibitor cocktail, by Roche (1 mentions) Complete ultra protease inhibitor, by Roche (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Jnk1 2, by Cell Signaling Technology (1 mentions) Phospho smad2 ser465 467, by Cell Signaling Technology (1 mentions) Phospho smad3 ser423 425, by Cell Signaling Technology (1 mentions) Phospho ampkα thr172, by Cell Signaling Technology (1 mentions) Nf κb p65, by Cell Signaling Technology (1 mentions) Phospho mtor ser2448, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

JNK1/2 (83 citations) Phospho-JNK1/2 (29 citations) Anti-phospho-JNK1/2 (Thr183/Tyr185, (2 citations)

12 protocols using phospho jnk1 2 thr183 tyr185

1

Probing JNK Signaling with ANS and Raman-labelled Compounds

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Cells were treated with ANS (Abcam) or Raman-labelled derivatives as indicated in the figure legend. Cells were washed with ice-cold PBS and subsequently lysed in RIPA buffer supplemented with cOmplete™ ULTRA protease inhibitor and PhosSTOP phosphatase inhibitor cocktails (Roche). Cleared lysates were resolved by SDS-PAGE. Primary antibodies used for Western blotting were as follows: phosphoJNK1/2 (Thr183/Tyr185; 1 : 1000), JNK1/2 (1 : 1000) and β-Actin (1 : 3000) (all Cell Signaling Technologies).
Tipping W.J., Lee M., Serrels A., Brunton V.G, & Hulme A.N. (2017). Imaging drug uptake by bioorthogonal stimulated Raman scattering microscopy †Electronic supplementary information (ESI) available: Fig. S1–S11, Table S1, Scheme S1, synthetic procedures and spectra for all labels and Raman-labelled anisomycin derivatives, drug uptake avi files. See DOI: 10.1039/c7sc01837a. Primary data files can be found at http://dx.doi.org/10.7488/ds/2046. Chemical Science, 8(8), 5606-5615.
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2

Comprehensive Western Blot Analysis

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Western blot analyses were performed as previously described (Jiang et al., 2016 (link); Jiang et al., 2017a ; Jiang et al., 2017b (link)). The HDAC Antibody Sampler Kit (#9928), Phospho-HDAC4Ser246/HDAC5Ser259/HDAC7Ser155 (#3443), HDAC2 (#5113), HDAC6 (#7558), H3K4me2 (#9725), H3K4me3 (#9727), H3K27me2 (#9728), H3K27me3 (#9733), Phospho-AKTSer473 (#4060), AKT (#4685), Phospho-mTORSer2448 (#5536), mTOR (#2983), Phospho-AMPKαThr172 (#2535), AMPKα (#5831), Phospho-MEK1/2Ser217/221 (#9154), Phospho-ERK1/2Thr202/Tyr204 (#4370), ERK1/2 (#4695), Phospho-JNK1/2 Thr183/Tyr185 (#4668), JNK1/2 (#9258), Phospho-p38Thr180/Tyr182 (#4511), p38 (#8690), Phospho-β-CateninSer675 (#4176), Non-phospho-β-CateninSer33/37/Thr41 (#8814), β-Catenin (#8480), Phospho-IKKα/βSer176/180 (#2697), IKKβ (#8943), Phospho-NF-κB p65Ser536 (#3033), NF-κB p65 (#8242), Phospho-Smad2Ser465/467 (#3108), Smad2 (#5339), Phospho-Smad3Ser423/425 (#9520), Smad3 (#9523), Phospho-Smad1/5Ser463/465 (#9516), and Smad1 (#6944) antibodies were obtained from Cell Signaling Technology. Antibodies against Histone H3 (ab1791), H3K36me2 (ab9049), H3K36me3 (ab9050), H4K20me3 (ab9053), H3K9me1 (ab9045), H3K9me2 (ab1220), H3K4me1 (ab8895), H3K23me1 (ab176132), H4K5ac (ab51997), H4K8ac (ab15823), H3K18ac (ab40888), H3K23ac (ab61234) and H4K12ac (ab46983) were purchased from Abcam.
Guo X., Fang Z.M., Wei X., Huo B., Yi X., Cheng C., Chen J., Zhu X.H., Bokha A.O, & Jiang D.S. (2019). HDAC6 is associated with the formation of aortic dissection in human. Molecular Medicine, 25, 10.
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3

Molecular Mechanisms of TSLP Regulation

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SSA and SSC were purchased from Sigma-Aldrich (St. Louis, MO, USA). TNF-α was obtained from ProSpec-Tany TechnoGene Ltd. (Ness Ziona, Israel). The firefly luciferase assay system was purchased from Promega (Madison, WI, USA). The web-based program MatInspector (http://www.genomatix.de/ (accessed on 17 December 2020)) was used for promoter analysis. Anti-TSLP antibody was obtained from Novus Biologicals (Centennial, CO, USA), and anti-EGR1, phospho-ERK1/2 (Thr202/Tyr204), phospho-p38 (Thr180/Tyr182), and phospho-JNK1/2 (Thr183/Tyr185) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-GAPDH antibody was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Secondary antibodies conjugated to Alexa Fluor 488 and rhodamine red-X were obtained from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). The MAPK, p38 kinase, and JNK inhibitors (U0126, SB203580, SP600125), DNCB, TB, and an H&E staining kit were purchased from Sigma-Aldrich.
Ahn S.S., Lee Y.H., Yeo H., Jung E., Lim Y, & Shin S.Y. (2022). Saikosaponin A and Saikosaponin C Reduce TNF-α-Induced TSLP Expression through Inhibition of MAPK-Mediated EGR1 Expression in HaCaT Keratinocytes. International Journal of Molecular Sciences, 23(9), 4857.
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4

Antibody Immunoblotting for Mitochondrial Proteins

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Antibodies against the following proteins were purchased from Santa Cruz Biotechnology (Dallas, TX): ANP (sc20158 1:200), β‐MHC (sc53090 1:200), TIM50 (sc55338 1:200), and ASK1 (sc7931, 1:200). The following antibodies were purchased from Cell Signaling Technology (Danvers, MA): GAPDH (#2118 1:1000), phospho‐ASK1Thr845 (3765, 1:1000), phospho‐MEK1/2Ser217/221 (#9154 1:1000), total‐MEK1/2 (#9122 1:1000), phospho‐ERK1/2Thr202/204 (#4370 1:1000), total‐ERK1/2 (#4695 1:1000), phospho‐JNK1/2Thr183/Tyr185 (#4668 1:1000), total‐JNK1/2 (#9258 1:1000), phospho‐P38Thr180/182 (#4511 1:1000), and total‐P38 (#9212 1:1000). Drp1 (#184272, 1:500), Mfn1 (#57602, 1:500), Mfn2 (#56889, 1:500) and Nrf2 (#31163, 1:500) were purchased from Abcam (Cambridge, MA), Fetal calf serum was purchased from HyClone (Waltham, MA). The other reagents for cell culture were purchased from Sigma (St. Louis, MO).
Tang K., Zhao Y., Li H., Zhu M., Li W., Liu W., Zhu G., Xu D., Peng W, & Xu Y. (2017). Translocase of Inner Membrane 50 Functions as a Novel Protective Regulator of Pathological Cardiac Hypertrophy. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(4), e004346.
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5

Antibody-Based Analysis of Neuronal Stress

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Mouse monoclonal antibodies against tyrosine phospho-Hydroxylase (Ser31), GAPDH, β-actin, 4-Hydroxy-2-nonenal (4-HNE), Nitro-tyrosine, CHOP, Fas, Bax, NFkB p65, Histone H1, TRAF2, IRE1α, and phospho-IKB-β were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit monoclonal antibodies against phospho-p38 MAPK (Thr180/Tyr182) and phospho-JNK1/2 (Thr183/Tyr185) were purchased from Cell Signaling Technology (Beverly, MA, USA). The TdT-mediated dUTP Nick End Labeling (TUNEL) kits were purchased from Roche (Germany). MPP+ (1-methyl-4-phenylpyridinium), SDS, NP-40, while sodium deoxycholate, protease inhibitor cocktail was purchased from Sigma (St. Louis, MO, USA).
Kuo H.C., Lu C.C., Shen C.H., Tung S.Y., Hsieh M.C., Lee K.C., Lee L.Y., Chen C.C., Teng C.C., Huang W.S., Chen T.C, & Lee K.F. (2016). Hericium erinaceus mycelium and its isolated erinacine A protection from MPTP-induced neurotoxicity through the ER stress, triggering an apoptosis cascade. Journal of Translational Medicine, 14, 78.
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6

Western Blot Analysis of Signaling Proteins

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Antibodies specific to the following proteins were purchased from Cell Signaling Technology (Danvers, MA, USA) and used in western blot experiments: phospho‐MEK1/2Ser32/36 (9154, 1:1000 dilution); total‐MEK1/2 (9122, 1:1000 dilution); phospho‐ERK1/2Thr202/Tyr204 (4370, 1:1000 dilution); total‐ERK1/2 (4695, 1:1000 dilution); phospho‐JNK1/2Thr183/Tyr185 (4668, 1:1000 dilution); total‐JNK (9258, 1:1000 dilution); phospho‐P38Thr180/Tyr182 (4511, 1:1000 dilution); and total‐P38 (9212, 1:1000 dilution). Antibodies specific to the following proteins were purchased from Santa Cruz Biotechnology (Dallas, TX): ANP (sc20158, 1:200 dilution) and β‐MHC (sc53090, 1:200 dilution). Anti‐GAPDH (MB001, 1:10000 dilution) was purchased from Bioworld Technology (Harrogate, UK). Anti‐ACE3 (BAM‐73‐006‐EX, 1:500 dilution) was purchased from COSMO BIO Co, Ltd (Tokyo, Japan). The BCA protein assay kit was purchased from Pierce (Rockford, IL). Peroxidase‐conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA) and used for visualization. Unless otherwise noted, cell culture reagents and all other reagents were obtained from Sigma (St. Louis, MO).
Yu C., Tang L., Liang C., Chen X., Song S., Ding X., Zhang K., Song B., Zhao D., Zhu X., Li H, & Zhang Z. (2016). Angiotensin‐Converting Enzyme 3 (ACE3) Protects Against Pressure Overload‐Induced Cardiac Hypertrophy. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(2), e002680.
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7

MAPK Signaling Pathway Protein Detection

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Cells or tissue samples were homogenized into lysis buffer containing 20 mM HEPES pH 7.4, 1% Triton X-100, 150 mM NaCl, 1 mM EDTA, and a protease/phosphatase inhibitor cocktail. Protein samples were quantified and subjected to 10% SDS-PAGE. The following antibodies were used for Western blot detection: phospho-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology, #9101), phospho-p38 (Thr180/Tyr182) (Cell Signaling Technology, #9211), phospho-JNK1/2 (Thr183/Tyr 185) (Cell Signaling Technology, #4668), phospho-MEK1/2 (Ser217/221) (Cell Signaling Technology, #9154), phospho-MKK3/MKK6 (Ser189/207) (Cell Signaling Technology, #9231), phospho-MKK7 (Ser271/Thr275) (Cell Signaling Technology, #4171), ERK1/2 (Cell Signaling Technology, #9102), p38 (Cell Signaling Technology, #9212), JNK (Cell Signaling Technology, #9252), MEK1/2 (Cell Signaling Technology, #4694), MKK7 (Cell Signaling Technology, #4172), MKK6 (Cell Signaling Technology, #9264), Flag (Cell Signaling Technology, #2368), α tubulin (Santa Cruz biotechnology, sc-5286), lamin A/C (Cell Signaling Technology, #2032), DUSP8 (Abcam, #ab184134), sarcomeric α-actinin (Sigma, #A7811), and GAPDH (Fitzgerald, #10R-G109A). Western blots were quantified using NIH ImageJ and normalized to total protein levels.
Liu R., van Berlo J.H., York A.J., Maillet M., Vagnozzi R.J, & Molkentin J.D. (2016). DUSP8 Regulates Cardiac Ventricular Remodeling by Altering ERK1/2 Signaling. Circulation research, 119(2), 249-260.
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8

Western Blot Analysis of Phospho-JNK1/2

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Cellular proteins were lysed, and equal amounts of protein (20 μg) were separated by SDS-PAGE and then transferred to PVDF membranes, which were incubated with primary antibodies, phospho-JNK1/2 (Thr183/Tyr185) and β-tubulin (Cell Signaling Technology, Danvers, MA; Abcam, Cambridge, UK). Bound antibody complexes were visualized using NBT/BCIP solution with color development buffer.
Jin X., Wang C., Wu W., Liu T., Ji B, & Zhou F. (2018). Cyanidin-3-glucoside Alleviates 4-Hydroxyhexenal-Induced NLRP3 Inflammasome Activation via JNK-c-Jun/AP-1 Pathway in Human Retinal Pigment Epithelial Cells. Journal of Immunology Research, 2018, 5604610.
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9

Protein Expression Analysis by Western Blot

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Protein concentrations were determined by Bradford assay. Albumin was used to establish the standard curve. Equal amounts of whole-cell lysates were subjected to 10% sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto PVDF membranes. Blots were incubated with primary Abs specific for phospho-p38 (T180/Y182, R&D Cell Signaling Technology, USA), phospho-ERK1/2 (Thr202/Tyr204, Cell Signaling Technology, USA), phospho-JNK1/2 (Thr183/Tyr185, Cell Signaling Technology, USA), NF-κB subunit p65 (Santa Cruz Biotechnology, USA), GAPDH (Jingmei Biotechnology, China) or proliferating cell nuclear antigen (PCNA, BD Biosciences, USA) at 25°C for 2 h. Subsequently, the bolts were incubated with peroxidase-conjugated secondary Abs (Boster Biotech) at 25°C for 1 h. The immunoreactive signals were visualized with chemiluminescence (Beyotime Institute of Biotechnology, China) according to the manufacturer's instructions and quantitated by Scion image software.
Fang L., Wang K.K., Huang Q., Cheng F., Huang F, & Liu W.W. (2020). Nucleolin Mediates LPS-induced Expression of Inflammatory Mediators and Activation of Signaling Pathways. Current medical science, 40(4).
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10

Cellular Signaling Pathway Modulation

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Dulbecco’s modified Eagle’s medium (DMEM)/F-12 medium, fetal bovine serum (FBS), and TRIzol were from Invitrogen (Carlsbad, CA, USA). The Hybond C membrane and enhanced chemiluminescence (ECL) Western blot detection system were from GE Healthcare Biosciences (Buckinghamshire, UK). The phospho-ERK1/2 (Thr202/Tyr204) (Cat.#4370), phospho-p38 (Thr180/Tyr182) (Cat.#4511), phospho-JNK1/2 (Thr183/Tyr185) (Cat.#4668), and phospho-p65 NF-κB (Ser536) (Cat.#3033) antibodies were from Cell Signaling (Danver, MA, USA). The anti-ZO-1 (Cat.#sc-33725) antibody was from Santa Cruz (Dallas, TX, USA). The anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from GeneTex (Irvine, CA, USA). The R-7050, U0126, SB202190, SP600125, Bay11-7082, and MMP2/9 inhibitor (2/9i) were from Enzo (Farmingdale, NY, USA). The bicinchoninic acid (BCA) protein assay reagent was from Pierce (Rockford, IL, USA). The tumor necrosis factor-α (TNF-α) was from R&D Systems (Minneapolis, MN, USA). The sophoraflavanone G (SG) was from ChemFaces (Wuhan, China). The enzymes and other chemicals were from Sigma (St. Louis, MO, USA).
Lee T.H., Chen J.L., Tsai M.M., Wu Y.H., Tseng H.C., Cheng L.C., Shanmugam V, & Hsieh H.L. (2023). Protective Effects of Sophoraflavanone G by Inhibiting TNF-α-Induced MMP-9-Mediated Events in Brain Microvascular Endothelial Cells. International Journal of Molecular Sciences, 25(1), 283.
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