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Total jnk1 2

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Total JNK1/2 is a lab equipment product that measures the total levels of c-Jun N-terminal kinases (JNK1 and JNK2) in cell lysates. It provides a quantitative assessment of the total JNK1 and JNK2 protein expression levels.

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Lab products found in correlation

Total erk1 2, by Cell Signaling Technology (5 mentions) Phospho jnk1 2, by Cell Signaling Technology (3 mentions) β actin, by Cell Signaling Technology (2 mentions) Total mek1 2, by Cell Signaling Technology (2 mentions) β mhc, by Santa Cruz Biotechnology (2 mentions) P jnk1 2, by Cell Signaling Technology (2 mentions) P erk1 2, by Cell Signaling Technology (2 mentions) P mtor, by Cell Signaling Technology (2 mentions) 2 7 dichlorofluorescin diacetate dcfda, by Merck Group (1 mentions) Total p38, by Cell Signaling Technology (1 mentions) Phospho jnk1 2 thr183 tyr185, by Cell Signaling Technology (1 mentions) Phospho p38thr180 182, by Cell Signaling Technology (1 mentions) Phospho ask1thr845, by Cell Signaling Technology (1 mentions) Phospho erk1 2thr202 204, by Cell Signaling Technology (1 mentions) Phospho mek1 2 ser217 221, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Total ampkα, by Cell Signaling Technology (1 mentions) P p70s6k, by Cell Signaling Technology (1 mentions) Thymoquinone tq, by Merck Group (1 mentions) Phenylephrine pe, by Tokyo Chemical Industry (1 mentions)

7 protocols using total jnk1 2

1

GHK-Cu Modulates LPS-Induced Inflammatory Responses

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GHK-Cu (MW 340.38; soluble in water) was provided by Bioceltran Corporation Ltd., (Chuncheon, Korea). Lipopolysaccharide (Escherichia coli O111:B4), thioclycollate and 2′,7′-dichlorofluorescin-diacetate (DCFDA) were purchased from Sigma (MO, USA). Antibodies against phospho-ERK1/2 (#4370), total ERK1/2 (#4695), phospho-p38 MAPK (#4511), total p38 MAPK (#9212), phospho-JNK1/2 (#4668), total JNK1/2 (#9252), phospho-NF-κB p65 (Ser536, #3033) and β-actin (#3700) were purchased from Cell Signaling (MA, USA). Antibodies against NF-κB p65 (sc-372) was purchased from Santa Cruz Biotechnology (CA, USA).
Park J.R., Lee H., Kim S.I, & Yang S.R. (2016). The tri-peptide GHK-Cu complex ameliorates lipopolysaccharide-induced acute lung injury in mice. Oncotarget, 7(36), 58405-58417.
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2

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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3

Signaling Pathway Antibody Analysis

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Antibodies against the following proteins were purchased from Cell Signaling Technology (USA): phosphorylated (p‐) AMPKα (Thr172) (used in experiments at 1:1,000 dilution), total AMPKα (at 1:1,000 dilution), p‐ACC (Ser79) (at 1:1,000 dilution), total ACC (at 1:1,000 dilution), p‐ERK1/2 (Thr202/Tyr204) (at 1:2,000 dilution), total ERK1/2 (at 1:2,000 dilution), p‐p38 (Thr180/Tyr182) (at 1:1,000 dilution), total p38 (at 1:1,000 dilution), p‐JNK1/2 (Thr183/Tyr185) (at 1:1,000 dilution), total JNK1/2(at 1:1,000 dilution), p‐mTOR (Ser2448) (at 1:1,000 dilution), total mTOR (at 1:1,000 dilution), p‐p70S6K (Thr389) (at 1:1,000 dilution), total p70S6K (at 1:1,000 dilution) and HRP‐linked secondary antibody (at 1:3,000 dilution). The antibody against GAPDH (at 1:3,000 dilution) was purchased from Abclonal. In addition, the following reagents were used: thymoquinone (TQ; Sigma‐Aldrich), phenylephrine (PE; Tokyo Chemical Industry) and compound C (CpC; Selleck).
Chen H., Zhuo C., Zu A., Yuan S., Zhang H., Zhao J, & Zheng L. (2021). Thymoquinone ameliorates pressure overload‐induced cardiac hypertrophy by activating the AMPK signalling pathway. Journal of Cellular and Molecular Medicine, 26(3), 855-867.
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4

Antibody and Kinase Inhibitor Protocolshop

Cited 1 time
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Antibodies against total ERK1/2 (#9102), phospho-ERK1/2 (Thr202/Tyr204; #9101), total IκBα (#9242), total JNK1/2 (#9252), phospho-p38MAPK (Thr180-Tyr182; #9211), total MK2 (#3042), phospho-MK2 (Thr334; #3042), total MSK1 (#3489), phospho-MSK1 (Thr581; #9595), phospho-c-Jun (Ser73; #9164) and phospho-IKKα/β (Ser176/180; #2697) were purchased from Cell Signaling Technology. Antibodies to Phospho-CREB (Ser133, #06-519) was from Millipore, anti-p38α (#sc-535) and anti-DUSP1 (#sc-373841) were from Santa Cruz, anti-active phospho-JNK1/2 (Thr183-Tyr185; #MAB1205) from R&D System, and anti-α-Tubulin (#T9026) from Sigma. Secondary antibodies from Invitrogen (Waltham, Massachusetts, United States) included Alexa Fluor 680 donkey α-sheep IgG (H + L) (#A21102), Alexa Fluor 680 goat α-rabbit IgG (H + L) (#A21109), Alexa Fluor 700 goat α-mouse IgG (H + L) (#A21036).
Kinase inhibitors SB203580 (inhibits p38α/p38β (Kuma et al., 2005 (link))) was purchased from Selleckchem, and SB747651A (inhibits MSK1, (Naqvi et al., 2012 (link))) from Axon Medchem. PD184352 (MKK1 inhibitor, (Kuma et al., 2005 (link))) and BIRB0796 (p38α/p38β/p38γ and p38δ inhibitor, (Kuma et al., 2005 (link))) were from the Division of Signal Transduction Therapy (DSTT); University of Dundee (Dundee, UK). JNK-IN-8 (JNK inhibitor (Zhang et al., 2012 (link))) was from Calbiochem.
Díaz-Mora E., González-Romero D., Meireles-da-Silva M., Sanz-Ezquerro J.J, & Cuenda A. (2023). p38δ controls Mitogen- and Stress-activated Kinase-1 (MSK1) function in response to toll-like receptor activation in macrophages. Frontiers in Cell and Developmental Biology, 11, 1083033.
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5

Platelet-Activating Factor Signaling Assay

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The following reagents were purchased from Calbiochem (Gibbstown, New Jersey, USA): Platelet-activating factor (PAF) was purchased from Biomol (Plymouth Meeting, Pennsylvania, USA). Antibodies (Ab) specific for phospho-ERK1/2, phospho-p38MAPK, phospho-JNK1/2, total-JNK1/2, total-ERK1/2, total-p38 MAPK and β-actin were purchased from Cell Signaling Technology (Danvers, MA, USA). All other reagents were purchased from Sigma Chemical Company (St. Louis, Missouri, USA). Anti-p47phox antibody was purchased from BD Biosciences (San Jose, California, USA) and anti-phospho-p47phox antibody was kindly donated by Dr. Jamel El-Benna. Phycoerythrin (PE)-conjugated anti-human CD63 monoclonal antibody (mAb) was purchased from Biolegend (San Diego, California, USA). All other reagents were purchased from Sigma Chemical Company unless otherwise stated.
Narantsogt G., Min A., Nam Y.H., Lee Y.A., Kim K.A., Agvaandaram G., Dorjsuren T., El-Benna J, & Shin M.H. (2015). Activation of MAPK Is Required for ROS Generation and Exocytosis in HMC-1 Cells Induced by Trichomonas vaginalis-Derived Secretory Products. The Korean Journal of Parasitology, 53(5), 597-603.
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6

Molecular Signaling Pathway Analysis

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Cholesterol and methyl-β-cyclodextrin were purchased from Sigma-Aldrich. Compound C, STO-609 acetate and CEP-1347 were purchased from Tocris Bioscience. Cytochalasin D, Jasplakinolide and Latrunculin B were purchased from Cayman Chemical. PD98059 was purchased from Enzo Life Sciences. SP600125 and U0126 were purchased from Calbiochem. Antibodies for phospho-AMPKα (#2535), MyD88 (#4283), phospho-ERK1/2 (#9101), total-ERK1/2 (#9102), phospho-JNK1/2 (#9251), total-JNK1/2 (#9252), anti-rabbit HRP-linked antibody (#7074) and anti-mouse HRP-linked antibody (#7076) were purchased from Cell Signaling Technology. Antibodies for AMPKα1 (sc-19128), MUC5AC (sc-21701), TAK1 (sc-7967), MLK3 (sc-166592), c-myc (sc-40), β-actin (sc-8432) and donkey anti-goat HRP-linked antibody (sc-2020) were purchased from Santa Cruz Biotechnology. Donkey anti-rabbit IgG Alexa Fluor 488 (A-21206) and goat anti-mouse IgG Alexa Fluor 546 (A-11030) was purchased from Thermo Fisher Scientific. The human validated short interfering RNA (siRNA) oligonucleotides for AMPKα1 (M-005027-02-0005), TAK1 (M-003790-06-0005) and control siRNA (D-001206-14-05) were purchased from Dharmacon.
Matsuyama S., Komatsu K., Lee B.C., Tasaki Y., Miyata M., Xu H., Shuto T., Kai H, & Li J.D. (2022). Negative cross-talk between TLR2/4-independent AMPKα1 and TLR2/4-dependent JNK regulates S. pneumoniae-induced mucosal innate immune response. Journal of immunology (Baltimore, Md. : 1950), 209(8), 1532-1544.
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7

Antibody Source and Reagent Details

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The antibodies against ANP (atrial natriuretic peptide) (sc20158) and β-MHC (β-myosin heavy chain) (sc53090) were from Santa Cruz Biotechnology. The antibody against TRIM32-(28–984) was from ProSci. The antibody against GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (MB001) was from Bioworld Technology. The antibodies against p-MEK1/2 [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase 1/2] (9154), total MEK1/2 (9122), p-ERK1/2 (4370), total ERK1/2 (4695), p-JNK1/2 (c-Jun N-terminal kinase 1/2) (4668), total JNK1/2 (9258), p-p38 (4511), total p38 (9212), p-Akt (4060), total Akt (4691), p-GSK3β (glycogen synthase kinase 3β) (9322), total GSK3β (9315), p-mTOR (mammalian target of rapamycin) (2971), total mTOR (2983) and total p70 (2708) were from Cell Signaling Technology. The antibody against p-p70 was from GeneTex (GTX50304). The BCA protein assay kit was from Pierce. FBS was from Hyclone. Cell culture reagents and all other reagents were from Sigma.
Chen L., Huang J., Ji Y., Zhang X., Wang P., Deng K., Jiang X., Ma G, & Li H. (2016). Tripartite motif 32 prevents pathological cardiac hypertrophy. Clinical Science (London, England : 1979), 130(10), 813-828.
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