Anti phospho c jun ser73

Manufactured by Cell Signaling Technology

Sourced in Japan, United States

Anti‐phospho‐c‐Jun (Ser73) is a primary antibody that detects the phosphorylated form of the c‐Jun protein at serine 73. It is used to analyze the activation of the c‐Jun transcription factor in cellular signaling pathways.

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Lab products found in correlation

Close spelling variants of this product

C-Jun (263 citations) Anti-c-Jun (99 citations) Phospho-c-Jun (97 citations) Anti-phospho-c-Jun (49 citations) Phospho-c-Jun (Ser73) (15 citations) Rabbit anti-phospho-c-Jun (Ser73) mAb (2 citations)

7 protocols using anti phospho c jun ser73

Immunoblotting Analysis of Kinase Signaling

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Protein samples (15 μg per lane) were subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes, and immunoblotted with anti‐MKK7, anti‐MKK4, anti‐JNK, anti‐phospho‐JNK, anti‐c‐Jun, anti‐phospho‐c‐Jun (Ser73), anti‐phospho‐histone H3 (Cell Signaling Technology), anti‐glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) (Cell Signaling Technology) and anti‐β‐actin (Sigma) antibodies. All blots were developed using an enhanced chemiluminescence detection system (GE Healthcare, Chalfont St. Giles, UK).

Ooshio T., Yamamoto M., Fujii K., Xin B., Watanabe K., Goto M., Okada Y., Suzuki A., Penninger J.M., Nishina H, & Nishikawa Y. (2021). Hepatocyte Mitogen‐Activated Protein Kinase Kinase 7 Contributes to Restoration of the Liver Parenchyma Following Injury in Mice. Hepatology (Baltimore, Md.), 73(6), 2510-2526.

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Granulosa Cell Culture Protocols

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All cell culture materials were obtained from Gibco Inc. Recombinant Human Activin-A (GFH6) was purchased from Cell Guidance Systems. RepSox (#72392), a cell permeable, selective inhibitor of the TGF-β type 1 receptor (TGFβRI) ALK5 was obtained from Stemcell Technologies. Recombinant forms of FSH (Gonal-F) and hCG (Ovitrelle) was purchased from Merck Global (Darmstadt, Germany). SAPK/JNK Kinase Assay Kit (#8794, nonradioactive), Hoechst 33342 (#4082), anti c-Jun (#9165), anti-phospho-c-Jun^Ser73 (#3270S), Anti-Phospho-SAPK/JNK^{Thr183/Tyr185} (#9251), Smad2 (#3122) and Phospho-Smad2^{Ser465/Ser467} (#18338) antibodies were obtained from Cell Signaling. Oil Red O was purchased from Sigma Inc. (USA). All western blotting buffers and reagents were purchased from Bio-Rad. Anti-Vinculin Antibody (V9264) was purchased from Sigma-Aldrich. Mouse antihuman monoclonal antibodies were purchased from Santa Cruz Biotechnology for the detection of human 3β-HSD Type II (sc-100466), 17β-HSD type-I (sc-376719), StAR (sc-166821), and P450 SCC (CYP11A1, sc-292456). Aromatase (CYP19A1, ab34193) monoclonal mouse antibody was from Abcam Inc. YO-PRO®-1 Iodide (491/509) was obtained from Life Technologies. Texas Red™-X Phalloidin was obtained from Thermo Fisher Scientific.

Bildik G., Akin N., Esmaeilian Y., Hela F., Yildiz C.S., Iltumur E., İncir S., Karahuseyinoglu S., Yakin K, & Oktem O. (2020). Terminal differentiation of human granulosa cells as luteinization is reversed by activin-A through silencing of Jnk pathway. Cell Death Discovery, 6, 93.

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Western Blot Protein Analysis

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Cell lysis, protein extraction, and western blot analyses were performed as described in our previous work (40 (link)). Proteins were dissolved in a lysis buffer and separated using SDS/PAGE for western blot analyses. Primary antibodies included rabbit anti-Phospho-SAPK/JNK (Thr183/Tyr185), anti-SAPK/JNK, anti-Phospho-c-Jun (Ser73), anti-c-Jun, anti-ATG5, anti-LC3B and anti-GAPDH (Cell Signaling Technology). Secondary antibody was HRP-conjugated anti-rabbit IgGs (Life Technologies). The densitometric analyses of western blotting images were performed using ImageJ software (National Institutes of Health).

Meng Q., Zhang Y, & Hu L.G. (2020). Targeting Autophagy Facilitates T Lymphocyte Migration by Inducing the Expression of CXCL10 in Gastric Cancer Cell Lines. Frontiers in Oncology, 10, 886.

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Histological and Immunohistochemical Analysis of Livers

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Livers were perfused with phosphate‐buffered saline and fixed in 4% paraformaldehyde overnight. Paraffin sections (4 μm) were deparaffinized and stained with hematoxylin and eosin (HE). Immunohistochemical analyses were performed by the Envision (peroxidase; Dako, Carpinteria, CA) method using anti‐Ki‐67 (Nichirei, Tokyo, Japan), anti–phospho‐c‐Jun (Ser73; Cell Signaling Technology, Danvers, MA), antifibrinogen (Abcam, Cambridge, UK), anti–α‐smooth muscle actin (α‐SMA; Dako), anti–cleaved caspase 3 (Cell Signaling Technology), anti–green fluorescent protein (GFP; ThermoFisher Scientific, Waltham, MA), antitransgelin (GeneTex, Irvine, CA), anti‐uPA (Abcam), and anti–plasminogen activator tissue‐type (tPA; Abcam) antibodies according to an antigen retrieval procedure using Target Retrieval Solution (Dako). To evaluate the extent of fibrosis, the sections were stained with sirius red F3B (Waldeck, Munster, Germany).

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Western Blot Analysis of Cell Signaling Proteins

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Whole-cell extracts were prepared using lysis buffer (Cell Signaling Technology). Cell lysates (20 μg) were separated by SDS-PAGE, and blotting performed as previously described.^{26 (link)} Rabbit anti-GSTA4 (Abnova; H00002941-D01P, 1:2,000), anti-Nrf2 (Cell Signaling Technology; #12721, 1:1,000), anti-phospho-c-Jun (Ser⁷³) (Cell Signaling Technology; #3270, 1:1,000), and anti-phospho-c-Fos (Ser³²) (Cell Signaling Technology; #5348, 1:1,000) were used as primary antibodies. Murine anti-β-actin loading control was purchased from Santa Cruz Biotechnology (sc-81178, 1:2,000). Goat anti-rabbit IgG-horseradish peroxidase (HRP) conjugate (Life Technologies; #65–6120, 1:10,000) and horse anti-mouse IgG-HRP conjugate (Cell Signaling Technology; #7076, 1:5,000) were used as secondary antibodies. Signals were generated by Clarity™ Western ECL Substrate (Bio-Rad). Images were captured by ChemiDoc XRS⁺ system and analyzed using Image Lab software (Bio-Rad).

Yang Y., Huycke M.M., Herman T.S, & Wang X. (2016). Glutathione S-transferase Alpha 4 Induction by Activator Protein 1 in Colorectal Cancer. Oncogene, 35(44), 5795-5806.

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Phospho-regulatory Pathway Profiling

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All cell culture materials were obtained from Gibco Inc. SP600125 and AS601245, pharmacological inhibitors of JNK, were purchased from Calbiochem. Control (#6568) and SAPK/JNK (#6232) siRNA, SAPK/JNK Kinase Assay Kit (#8794, nonradioactive), Hoechst 33342 (#4082), Anti-c-Jun (#9165), Anti-Phospho-c-Jun^Ser63 (#12598), Anti-Phospho-c-Jun^Ser73 (#3270 S), Anti SAPK/JNK (#9252), Anti-Phospho-SAPK/JNK^{Thr183/Tyr185} (#9251), Anti-Cyclin A (4656), Anti-Cyclin B1 (#12231) and Anti-Phospho-cdc-2^Tyr15 (#4539) antibodies were obtained from Cell Signaling. All western blotting buffers and reagents were purchased from Bio-Rad. Anti-phospho-Histone H3^Ser10 (06-570) antibody was obtained from Upstate. Anti-Vinculin Antibody (V9264) was purchased from Sigma-Aldrich. TransIT-X2® Dynamic Delivery System (MIR 6000) was purchased from Mirus Bio LLC. YO-PRO®-1 Iodide (491/509) was obtained from Life Technologies. Texas Red™-X Phalloidin and Click-iT™ EdU Alexa Fluor™ 488 Imaging Kit (C10337) were obtained from Thermo Fisher Scientific. Matrigel Basement Membrane Matrix (356230) was from BD Biosciences. Super Block reagent (#AAA125) was purchased from ScyTek Laboratories.

Bildik G., Akin N., Senbabaoglu F., Esmalian Y., Sahin G.N., Urman D., Karahuseyinoglu S., Ince U., Palaoglu E., Taskiran C., Arvas M., Guzel Y., Yakin K, & Oktem O. (2018). Endogenous c-Jun N-terminal kinase (JNK) activity marks the boundary between normal and malignant granulosa cells. Cell Death & Disease, 9(4), 421.

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Antibody-based Protein Analysis in Autophagy

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Antibodies against microtubule-associated protein light chain 3 (LC3) and α-tubulin were obtained from Sigma Chemical Co. (St Louis, MO, USA) and Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA), respectively. Anti-p21^Waf1/Cip1, anti-p27^Kip1, anti-cyclin D1, anti-caspase-3, poly (ADP-ribose) polymerase (PARP), anti-c-jun-N-terminal kinase (JNK), anti-phospho-JNK (Thr183/Tyr185), anti-adenosine monophosphate-activated protein kinase (AMPK), anti-phospho-AMPK (Thr172), anti-ULK, anti-phospho-ULK (Ser555), anti-c-jun, anti-phospho-c-jun (Ser73), anti-ERK1/2, anti-phospho-ERK1/2 (Thr202)/Tyr204), anti-p38, anti-phospho-p38 (Thr180/Tyr182), anti-p53, anti-phospho-p53 (Ser15), anti-Bcl-2, anti-p62/SQSTM1, and anti-Beclin-1 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Propidium iodide (PI), Ribonuclease A (RNase A) from bovine pancreas, monodansyl cadaverine (MDC), 2′7′-dichlorofulorescein diacetate (DCF-DA), Bafilomycin A1 from Streptomyces griseus, and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma Chemical. SP600125, SB203580, PD98059, and N-acetyl-L-cysteine (NAC) were obtained from Calbiochem (San Diego, CA).

Kim A., Im M., Yim N.H., Kim T, & Ma J.Y. (2014). A Novel Herbal Medicine, KIOM-C, Induces Autophagic and Apoptotic Cell Death Mediated by Activation of JNK and Reactive Oxygen Species in HT1080 Human Fibrosarcoma Cells. PLoS ONE, 9(5), e98703.

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