Anti jnk

Manufactured by Santa Cruz Biotechnology

Sourced in United States, Germany

Anti-JNK is a laboratory reagent used for the detection and quantification of the JNK (c-Jun N-terminal kinase) protein. JNK is a member of the mitogen-activated protein kinase (MAPK) family and plays a crucial role in cellular signal transduction pathways. Anti-JNK is a specific antibody that can be used in various immunoassay techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to identify and measure the expression levels of JNK in biological samples.

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Close spelling variants of this product

Anti-p-JNK (50 citations) Anti-phospho-JNK (22 citations) Mouse anti-p-JNK (6 citations) Anti-JNK antibodies (5 citations) Anti-p-JNK (sc-6254) (4 citations) Rabbit anti-JNK (4 citations) Mouse anti-JNK (3 citations) Rabbit anti‐JNK (sc‐7345 (2 citations) Mouse monoclonal anti-JNK (2 citations) Anti-JNK and anti-p38 antibodies (2 citations)

71 protocols using anti jnk

Western Blot Analysis of Neurological Signaling

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After washing with ice-cold PBS, cells were lysed with RIPA lysis buffer (Thermo Fisher) containing protease inhibitor (Roche). The protein concentration in the lysates was measured using a NanoDrop spectrophotometer (ND-1000, Thermo Fisher), loading 20 μg of sample for each analysis. After separation in a 10% SDS-PAGE gel, proteins were transferred to polyvinylidene fluoride (PVDF) membranes, which were blocked with 5% skim milk in Tris-buffered saline plus Tween (TBST). The blots were incubated at 4 °C overnight with the following primary antibodies: anti-LINGO-1 (1:500, Abcam), anti-Wnt5a (1:250, R&D Systems), anti-JNK (1:800, Santa Cruz), anti-p-JNK (1:800, Santa Cruz), anti-β-catenin (1:1000, Cell Signaling Technology), and anti-β-actin (1:2000, Cell Signaling Technology). After washing with TBST, the membranes were incubated for 1 h with the appropriate secondary antibody (1:2000, Abcam). The developed X-ray films were scanned and images were analyzed quantitatively with ImageJ software (version 1.45, NIH, USA). Relative protein levels were expressed as fold change after normalization to β-actin.

Zhao C.G., Qin J., Li J., Jiang S., Ju F., Sun W., Ren Z., Ji Y.Q., Wang R., Sun X.L., Mou X, & Yuan H. (2021). LINGO-1 regulates Wnt5a signaling during neural stem and progenitor cell differentiation by modulating miR-15b-3p levels. Stem Cell Research & Therapy, 12, 372.

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Immunoblotting Analysis of Protein Signaling

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The anti-TDP-43 antibody was described previously,^{66 (link)} and the following additional primary antibodies were used: anti-iNOS (Abcam, Cambridge, MA, USA), anti-COX-2 (Epitomics, Burlingame, CA, USA), anti-GAPDH (Chemicon, Temecula, CA, USA), anti-phospho-MEK (S217/221) (Cell Signaling Technology, Beverly, MA, USA), anti-MEK (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-phospho-ERK (Santa Cruz Biotechnology), anti-ERK (Santa Cruz Biotechnology), anti-phospho-JNK (Epitomics), anti-JNK (Santa Cruz Biotechnology), anti-phospho-p38 (Epitomics), anti-p38 (Santa Cruz Biotechnology), anti-phospho-c-Raf (S338) (Cell Signaling Technology), anti-c-JUN (Proteintech, Chicago, IL, USA), anti-phospho-Glycogen Synthase (S641) (p-GS) (Epitomics) and anti-MAP2 (Santa Cruz Biotechnology). The secondary antibodies used included horseradish peroxidase-conjugated sheep anti-mouse and anti-rabbit antibodies (Amersham Pharmacia Biotech, Peapack, NJ, USA). The antibody-bound proteins were visualized using an ECL detection kit (Amersham Biosciences, Piscataway, NJ, USA) or Alexa Fluor 488 (green) donkey anti-mouse IgG (Invitrogen, La Jolla, CA, USA).

Xia Q., Hu Q., Wang H., Yang H., Gao F., Ren H., Chen D., Fu C., Zheng L., Zhen X., Ying Z, & Wang G. (2015). Induction of COX-2-PGE2 synthesis by activation of the MAPK/ERK pathway contributes to neuronal death triggered by TDP-43-depleted microglia. Cell Death & Disease, 6(3), e1702-.

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Protein Expression Analysis by Immunoblotting

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Protein analysis was performed using standard immune blotting. The following antibodies were used at the indicated dilution: anti-Noxa (SC-2697) 1:1000; anti-cytochrome (#4212), 1:1000; anti-caspase 3 (#7190), 1:1000; anti-caspase 9 (#9501), 1:1000; anti-PARP (#9542), 1:500 (each Cell Signaling Technology Inc., Danvers, MA, USA); anti-IDO antibody 1:500 (BioGenes, Berlin, Germany); anti-ASK1 (Sc-7931), 1:500; anti-p-ASK1 (Sc-109911), 1:1000; anti-JNK (Sc-474), 1:1000; anti-p-JNK (SC-6254), 1:1000; anti-p38 (Sc-535), 1:1000; anti-p-p38 (Sc-7973), 1:1000; anti-Actin (Sc-1615), 1:5.000; anti-Tom20 (Sc-11415), 1:100; anti-Bap31 (Sc-18579), 1:1000; anti-HO-1 (sc-10789), 1:1000; anti-p-JAK1 (sc-16773), 1:1000, anti-IRE1α (Sc-20790), 1:500; anti-PERK (SC-9477), 1:1000; (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); anti-IκBα (Sc-7182), 1:1000; anti-p-IκBα (AF4809, R&D system), 1:1000; anti-JAK1 antibody (ab47435), 1:1000; anti-IRF1 antibody (ab55330), 1:1000 (each ABCAM).

El Jamal S.M., Taylor E.B., Abd Elmageed Z.Y., Alamodi A.A., Selimovic D., Alkhateeb A., Hannig M., Hassan S.Y., Santourlidis S., Friedlander P.L., Haikel Y., Vijaykumar S., Kandil E, & Hassan M. (2016). Interferon gamma-induced apoptosis of head and neck squamous cell carcinoma is connected to indoleamine-2,3-dioxygenase via mitochondrial and ER stress-associated pathways. Cell Division, 11, 11.

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Protein Expression Analysis Protocol

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The cells were lysed in RIPA with 1 mM PMSF and protease inhibitors (Sigma) for 15 min on ice. The cytosol and nuclear fraction were isolated according to the manufactory's instruction (Thermo). Similar amounts of protein from each extract were subjected to SDS-PAGE analysis and transferred to polyvinyl difluoride (PVDF) membranes (Millipore). After blocking for 1 h with blocking buffer (5% nonfat milk and 0.1% Tween-20 in PBS), the membranes were incubated with the following primary antibodies at 4°C overnight: anti-phospho-p38 MAPK, anti-p38 MAPK, anti-phospho-JNK, anti-JNK (Santa Cruz), anti-phospho-ERK, anti-ERK, anti-phospho-Akt, anti-Akt, anti-phospho-IκBα, anti-IκBα, anti-p50/p105, anti-p65, anti-Histone H3 (Cell Signaling Technology), anti-gC1qR (Hycult biotech) and anti-β-actin (Tianjin Sungene Biotech) antibodies. HRP-conjugated anti-mouse IgG or anti-rabbit IgG (Santa Cruz) were used as secondary antibodies. And ECL (Bio-Rad) was used for chemiluminescent detection according to the manufacturer's instructions.

Du Q., Huang Y., Wang T., Zhang X., Chen Y., Cui B., Li D., Zhao X., Zhang W., Chang L, & Tong D. (2016). Porcine circovirus type 2 activates PI3K/Akt and p38 MAPK pathways to promote interleukin-10 production in macrophages via Cap interaction of gC1qR. Oncotarget, 7(14), 17492-17507.

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Assessing Synthetic Amylin Internalization

Cited 1 time

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Antioxidant N-acetyl cysteine (NAC) (Sigma, MO) was used at a working concentration of 5 mM. Ask1 inhibitor NQDI1 (R&D Systems) and JNK inhibitor (JK1) (Abcam) were used at a working concentration of 10 µM. Lactacystin (Lac) (Calbiochem, CA) was used at a concentration of 10 µM. H₂O₂ (Sigma) and diamide (Sigma) were used at working concentrations of 500 µM and 1mM respectively. Anti-PARP (rabbit, sc-7150), anti-actin (goat, sc-1616), anti-ASK1 (goat, sc-6368), anti-phospho-ASK1 (rabbit, sc-1099), anti-caspase-3 (goat, sc-1224), anti-JNK (rabbit, sc-572) and anti-phospo-JNK (goat, sc-12882) polyclonal primary antibodies were purchased from Santa Cruz Biotechnology. Insulin antibody (sc-9168) was purchased from Santa Cruz Biotech. For detection of overexpressed human amylin in transgenic mice islets, mouse monoclonal anti-human antibody (Santa Cruz Biotech., sc-377530) was used. For detection of internalized synthetic human amylin, rabbit polyclonal anti-amylin antibody (Santa Cruz Biotech., CA sc-20936) was used. While both anti-amylin antibodies show high binding affinity toward pro- and pre/pro-human amylin isoforms, only polyclonal anti-hA antibody detects synthetic human amylin (Figure S1A), which was used in amylin internalization studies (Fig. 1C) [41 (link)].

Singh S., Bhowmick D.C., Pany S., Joe M., Zaghlula N, & Jeremic A.M. (2018). Apoptosis Signal Regulating Kinase-1 and NADPH Oxidase Mediate Human Amylin Evoked Redox Stress and Apoptosis in Pancreatic Beta-Cells. Biochimica et biophysica acta. Biomembranes, 1860(9), 1721-1733.

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Boehmeria nivea Extract Inhibits Mast Cell Activation

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Anti-DNP IgE, DNP-bovine serum albumin (BSA), dexamethasone, and Evans blue were purchased from Sigma (St. Louis, MO, USA). The RBL-2H3 cell line (KCLB-22256) was purchased from the Korean Cell Line Bank (Seoul, Korea). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, and penicillin-streptomycin (PS) were purchased from Hyclone (Logan, UT, USA). The histamine, TNF-α, IL-6, IL-1β, and IL-4 ELISA kits were purchased from Biovision (Milpitas, CA, USA). Anti-phospho-p38 (Thr180/Tyr182), anti-phospho-ERK (L352), anti-phospho-JNK (T183/Y185), anti-p38, anti-ERK, anti-JNK, and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Boehmeria nivea is an eco-friendly pesticide-free product, which was purchased from a farming association (Yeonggwang, Korea).

Lim J.Y., Lee J.H., Lee B.R., Kim M.A., Lee Y.M., Kim D.K, & Choi J.K. (2020). Extract of Boehmeria nivea Suppresses Mast Cell-Mediated Allergic Inflammation by Inhibiting Mitogen-Activated Protein Kinase and Nuclear Factor-κB. Molecules, 25(18), 4178.

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Comprehensive Immunoblotting Antibody Panel

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Anti-PKR (sc-708, Santa Cruz), anti-peIF2alpha (44728G, Invitrogen), anti-eIF2alpha (#9772, Cell Signaling), anti-pJNK (#4668, Cell Signaling), anti-JNK (sc-7345, Santa Cruz), anti-pIRS1 (07–247, Millipore), anti-IRS1 (sc-559, Santa Cruz), anti-pIKKbeta (#2694, Cell Signaling), anti-IKKbeta (#2370, Cell Signaling), and anti-Tubulin (T9026, Sigma).

Taga M., Mouton-Liger F., Sadoune M., Gourmaud S., Norman J., Tible M., Thomasseau S., Paquet C., Nicoll J.A., Boche D, & Hugon J. (2018). PKR modulates abnormal brain signaling in experimental obesity. PLoS ONE, 13(5), e0196983.

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Western Blot Analysis of Lung Proteins

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Lung samples were homogenized in ice-cold RIPA buffer (50 mmol/L Tris, 150 mmol/L NaCl, 1% Triton X-100, pH 7.4) containing protease inhibitor cocktail (Roche Life Science Co., USA). The whole cell lysates were obtained by centrifuging the suspensions for 10 min at 13,000 ×g at 4°C. The lysates were loaded onto a 10% SDS-polyacrylamide gel for separation and then transferred to nitrocellulose membranes. The blots were probed with anti-p-Jun N-terminal kinase (JNK) (1:400; Santa Cruz Biotechnology, Inc., USA), anti-JNK (1:400; Santa Cruz Biotechnology, Inc., USA), and anti-β-actin (1:400; Santa Cruz Biotechnology, Inc., USA) antibodies. After incubation with horseradish peroxidase-linked secondary antibodies, specific protein bands on the blots were visualized with the ECL chemiluminescence detection system (Millipore, USA) according to the manufacturer's manual.

Tian W.F., Weng P., Sheng Q., Chen J.L., Zhang P., Zhang J.R., Du B., Wu M.C., Pang Q.F, & Chu J.J. (2017). Biliverdin Protects the Isolated Rat Lungs from Ischemia-reperfusion Injury via Antioxidative, Anti-inflammatory and Anti-apoptotic Effects. Chinese Medical Journal, 130(7), 859-865.

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Evaluation of Endothelial Cell Activation Markers

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We obtained endothelial cell basal medium-2 (EGM-2) from Lonza Cambrex (Nottingham, UK) and obtained M-199 and RPMI 1640 tissue culture mediums from GIBCO (Grand Island, NY, USA). We obtained the anti-ICAM-1, anti-RAGE, anti-ERK, anti-phosphorylated-ERK, anti-JNK, anti-phosphorylated-JNK, anti-phosphorylated-p38, anti-NF-κB and anti-PCNA antibodies from Santa Cruz Biotechnology, Inc. (Heidelberg, Germany) and the antibodies against anti-VCAM-1, anti-p38, and phosphorylated-IkappaBα, PD98059 (an ERK1/2 inhibitor), SP600125 (a JNK inhibitor), and SB203580 (a p38 inhibitor) from Cell Signaling Technology Inc. (Danvers, MA, USA). We obtained the anti-GAPDH antibody from Merck (Darmstadt, Germany), and all the other most highly purified chemicals were commercially provided from Sigma-Aldrich (St. Louis, MO, USA).

Son W.R., Nam M.H., Hong C.O., Kim Y, & Lee K.W. (2017). Plantamajoside from Plantago asiatica modulates human umbilical vein endothelial cell dysfunction by glyceraldehyde-induced AGEs via MAPK/NF-κB. BMC Complementary and Alternative Medicine, 17, 66.

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Molecular Signaling Pathways in Inflammation

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Anti-phospho-JNK, anti-phospho-ERK and anti-phospho-p38 MAPK antibodies were obtained from Cell Signaling Technology (Beverly, MA). Anti-JNK, anti-ERK and anti-p38 MAPK antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-iNOS antibody was from eBioscience (San Diego, CA). Anti-β-actin antibody was from Sigma (St. Louis, MO). NF-κB inhibitor (JSH-23), JNK inhibitor (SP600125) and p38 MAPK inhibitor (SB 203580) were from Merck (Darmstadt, Germany). JAK inhibitor (Ruxolitinib) was obtained from Selleck Chemicals (Houston, TX). iNOS inhibitor (L-NMMA) was from Sigma (St. Louis, MO). Recombinant mouse IL-1β was obtained from PeproTech (Rocky Hill, NJ). Mouse IL-1β neutralizing antibody was from R&D Systems (Minneapolis, MN). Middlebrook 7H9 broth medium and Middlebrook 7H10 agar were purchased from BD Difco Laboratories (Sparks, MD).

Yang K., Wu Y., Xie H., Li M., Ming S., Li L., Li M., Wu M., Gong S, & Huang X. (2016). Macrophage-mediated inflammatory response decreases mycobacterial survival in mouse MSCs by augmenting NO production. Scientific Reports, 6, 27326.

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