Sapk jnk antibody

Manufactured by Cell Signaling Technology

Sourced in United States

The SAPK/JNK Antibody is a laboratory reagent used to detect and study the SAPK/JNK (Stress-Activated Protein Kinase/c-Jun N-Terminal Kinase) protein in various biological samples. It is a highly specific and sensitive antibody that recognizes the SAPK/JNK protein, which is involved in cellular stress response and signal transduction pathways.

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Close spelling variants of this product

SAPK/JNK (140 citations) JNK antibody (10 citations) Phospho-SAPK/JNK antibody (4 citations) Anti-SAPK/JNK antibody (3 citations) Anti-Phospho-SAPK/JNK (Thr183/Tyr185) antibody (3 citations) Rabbit anti-SAPK/JNK antibody (2 citations) Mouse anti-phospho-SAPK/JNK antibody (2 citations) Anti-phospho-SAPK/JNK (Thr183/Tyr185) rabbit polyclonal antibody (2 citations) Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (2 citations)

12 protocols using sapk jnk antibody

Western Blot Analysis of MAPK Signaling

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Western blot studies were performed as previously described^{6 (link)}. After the LPS and LECT2 stimulation, treated cells were collected and lysed in 1 × RIPA lysis buffer (Upstate Biotechnology) with a Complete Mini EDTA-free cocktail tablet (Roche Diagnostics) and PhosSTOP phosphatase inhibitor cocktail tablets (Roche Diagnostics). Protein samples were subjected to SDS-PAGE and transferred to polyvinylidene fluoride membranes, using an iBlot gel transfer system (Invitrogen). Membranes were blocked and then they were probed with antibodies for 16 h (Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (Cell Signaling, Danvers, MA), SAPK/JNK Antibody (Cell Signaling, Danvers, MA), Phospho-p38 MAPK (Thr180/Tyr182) Antibody (Cell Signaling, Danvers, MA), p38 MAPK Antibody (Cell Signaling, Danvers, MA), Phospho-p44/42 MAPK(Erk1/2) (Thr202/Tyr204) Antibody (Cell Signaling, Danvers, MA), p44/42 MAPK(Erk1/2) Antibody (Cell Signaling, Danvers, MA) Phospho-SEK1/MKK4 (Ser257) Antibody (Cell Signaling, Danvers, MA), SEK1/MKK4 Antibody (Cell Signaling, Danvers, MA) Afterward, membranes were washed and then incubated with anti-rabbit IgG horseradish peroxidase-linked antibody (Cell Signaling) for 1 h. Protein bands were visualized with ECL Prime Western blotting detection reagent (GE Healthcare UK Ltd.).

Takata N., Ishii K.A., Takayama H., Nagashimada M., Kamoshita K., Tanaka T., Kikuchi A., Takeshita Y., Matsumoto Y., Ota T., Yamamoto Y., Yamagoe S., Seki A., Sakai Y., Kaneko S, & Takamura T. (2021). LECT2 as a hepatokine links liver steatosis to inflammation via activating tissue macrophages in NASH. Scientific Reports, 11, 555.

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Osteoblast and Osteoclast Culture Techniques

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In vitro primary osteoblast cultures, osteoclast cultures, quantitative PCR, Western blotting, ALP and Von Kossa were performed as previously described.^{(11 (link))} Primary antibodies were specific for phospho-JUN, (1:1,000; Cell Signaling) and GAPDH (1:2,000; Sigma), Phospho-SAPK/JNK (Thr183/Tyr185), and SAPK/JNK Antibody (#9251 and #9252 Cell Signaling). Osteoclast culture and RNA isolation from whole bone was performed as previously described.^{(13 (link))} Osteocyte expression analysis was performed in the IDG-SW3 cell line, a generous gift form Dr. Lynda Bonewald.^{(12 (link))}

Xu R., Zhang C., Shin D.Y., Kim J.M., Lalani S., Li N., Yang Y.S., Liu Y., Eiseman M., Davis R.J., Shim J.H, & Greenblatt M.B. (2017). c-Jun N-terminal kinases (JNKs) are critical mediators of osteoblast activity in vivo. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 32(9), 1811-1815.

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MAPK Signaling Pathway Antibodies

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Rabbit anti-LRCH1 antibody (bs-9327R) was purchased from Beijing Bioss Antibodies Inc. The following antibodies were purchased from Cell Signaling Technology: beta-actin antibody (3700), phospho-p38 mitogen-activated protein kinase (MAPK) (Thr180/Tyr182) antibody (9216), phospho-SAPK/Jun N-terminal kinase (JNK) (Thr183/Tyr185) antibody (9255), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (9101), p44/42 MAPK (Erk1/2) antibody (4695), p38 MAPK antibody (8690), and SAPK/JNK Antibody (9252).

Chen W.K., Feng L.J., Liu Q.D., Ke Q.F., Cai P.Y., Zhang P.R., Cai L.Q., Huang N.L, & Lin W.P. (2020). Inhibition of leucine-rich repeats and calponin homology domain containing 1 accelerates microglia-mediated neuroinflammation in a rat traumatic spinal cord injury model. Journal of Neuroinflammation, 17, 202.

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Protein Extraction and Western Blot Analysis

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Total protein was extracted using a combination of RIPA buffer, phenylmethylsulfonyl fluoride, and phosphatase inhibitors. The protein lysates were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then transferred to poly(vinylidene fluoride) membranes. The membranes were blocked in 5% skim milk at room temperature for 1 h and then incubated with the designated primary antibodies at 4°C overnight. The excess primary antibody was washed off with Tris-buffered saline with Tween 20, and the corresponding secondary antibody was incubated at room temperature for 1 h. A United Kingdom UV imaging system was used for exposure imaging. ImageJ was used to analyze the experimental results. Western Blot antibodies:ATM antibody, Cell signaling (#2873); Phospho-ATM (Ser1981) antibody, Cell signaling (#13050); LC3B antibody, Abcam (AB192890); SAPK/JNK Antibody, Cell signaling (#9252); Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) antibody (#4668); AMBRA1 antibody, AB clonal (A12578); GAPDH antibody, AB clonal (AC002).

Wang Q., Chen Y., Chang H., Hu T., Wang J., Xie Y, & Cheng J. (2021). The Role and Mechanism of ATM-Mediated Autophagy in the Transition From Hyper-Radiosensitivity to Induced Radioresistance in Lung Cancer Under Low-Dose Radiation. Frontiers in Cell and Developmental Biology, 9, 650819.

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Anti-inflammatory Effects of Lomerizine

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Lomerizine was purchased from Absin Bioscience Inc. (Shanghai, China). Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St. Louis, United States). Dexamethasone was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Hematoxylin and eosin (H&E) staining kit was purchased from Abcam (Waltham, Boston, United States). Dulbecco’s Modified Eagle Medium (DMEM) and macrophage colony-stimulating factor (M-CSF) was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Penicillin-streptomycin solution (Pen Strep) was purchased from Yeasen Biotechnology Co., Ltd. (Shanghai, China). Fetal Bovine Serum (FBS) was purchased from Cegrogen Biotech (Wupperweg, Germany).
Primers were synthesized by Hua Gene Biotech Co., Ltd. (Shanghai, China). The primary antibody against IκB-α was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States). Primary antibodies against iNOS, phospho-NF-κB p65 (Ser536) Rabbit mAb, NF-kappaB p65 Rabbit mAb, phospho-p38 MAPK (Thr180/Tyr182) XP Rabbit mAb, p38 MAPK XP Rabbit mAb, phospho-SAPK/JNK (Thr183/Tyr185) Rabbit mAb, SAPK/JNK Antibody, Phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) Antibody, p44/42 MAPK (ERK1/2) Rabbit mAb, and β-Actin Rabbit Antibody were purchased from Cell Signaling Technology (Danvers, MA, United States).

Song Y., Gou Y., Gao J., Chen D., Zhang H., Zhao W., Qian F., Xu A, & Shen Y. (2023). Lomerizine attenuates LPS-induced acute lung injury by inhibiting the macrophage activation through reducing Ca2+ influx. Frontiers in Pharmacology, 14, 1236469.

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Antibody-Mediated Signaling Pathway Analysis

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HH cells were cultured in six-well plates in the presence or absence of anti-human CD147 antibody (1 µg/mL; Biolegend) or anti-human CypA antibody (0.7 µg/mL; Abcam) for 1, 3, or 6 h. After collecting proteins from HH cells, equal amounts of proteins were subjected to 4–12% NuPage Bis-Tris Gels (Invitrogen, Waltham, MA, USA) at 200 V for 20 min. The proteins were then transferred onto polyvinylidene fluoride membranes (Invitrogen) and blocked in 2% skim milk powder with 0.05% Tween-20 (Sigma-Aldrich, St. Louis, MO, USA) in Tris-buffered saline. The membranes were probed with Akt Antibody (Cell Signaling Technology, Danvers, MA, USA), Phospho-Akt Antibody (Cell Signaling Technology), ERK1/2 Antibody (Cell Signaling Technology), Phospho-ERK1/2 Antibody (Cell Signaling Technology), p38 MAPK Antibody (Cell Signaling Technology), Phospho-p38 MAPK Antibody (Cell Signaling Technology), SAPK/JNK Antibody (Cell Signaling Technology), Phospho-SAPK/JNK Antibody (Cell Signaling Technology), or β-actin Antibody (Cell Signaling Technology) as primary antibody overnight at 4 °C, followed by incubation in secondary antibody for 30 min at room temperature. Visualization was performed by SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, Waltham, MA, USA).

Sakamoto M., Miyagaki T., Kamijo H., Oka T., Boki H., Takahashi-Shishido N., Suga H., Sugaya M, & Sato S. (2021). CD147-Cyclophilin a Interactions Promote Proliferation and Survival of Cutaneous T-Cell Lymphoma. International Journal of Molecular Sciences, 22(15), 7889.

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Molecular Signaling Pathway Analysis

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Galectin‐3 antibody, SAPK/JNK antibody, phospho‐SAPK/JNK (Thr183/Tyr185) antibody, Phospho‐PI3 Kinase Class III (Ser249) antibody, PI3 Kinase Class III antibody, SQSTM1/p62 antibody, LC3B antibody, and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) antibody were purchased from Cell Signaling (Danvers, MA, USA). Collagen type 1A1 (COL1A1) antibody and interleukin (IL)‐1R‐associated kinase 1 (IRAK‐1) were purchased from Santa Cruz (Dallas, TX, USA). The α‐SMA antibody was purchased from Sigma‐Aldrich (St. Louis, MO, USA). Alexa Fluor 488 goat anti‐mouse immunoglobulin G (IgG) and Alexa Fluor 594 goat anti‐rabbit immunoglobulin G were purchased from Invitrogen (Carlsbad, CA, USA). DOX hydrochloride and SP600125 were purchased from Sigma‐Aldrich. Chloroquine was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Pioglitazone hydrochloride was purchased from Fujifilm (Tokyo, Japan). ODN2088 and GW1929 were purchased from AdipoGen Life Sciences (San Diego, CA, USA).

Tanaka R., Umemura M., Narikawa M., Hikichi M., Osaw K., Fujita T., Yokoyama U., Ishigami T., Tamura K, & Ishikawa Y. (2020). Reactive fibrosis precedes doxorubicin‐induced heart failure through sterile inflammation. ESC Heart Failure, 7(2), 588-603.

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Cellular Signaling Pathway Analysis

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High glucose Dulbecco's Modified Eagle's Medium (#11995065), fetal bovine serum (#10270) were purchased from Thermo Fisher Scientific (Gibco, Waltham, MA, USA). Thioglycolate medium Brewer (Lot #5243569) was purchased from Becton, Dickinson and Company (Sparks, MD, USA). RIPA Buffer (#R0020) was purchased from Solarbio (Beijing, China). Antibodies were as follows: Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb (#4668), SAPK/JNK Antibody (#9252), β-Actin (8H10D10) Mouse mAb (#3700), Anti-rabbit IgG HRP-linked Antibody (#7074) and Anti-mouse IgG HRP-linked Antibody (#7076) were purchased from Cell Signaling Technology (Boston, MA, USA); Phospho-c-Jun-S63 Rabbit mAb (AP0105), JUN Rabbit pAb (#A16905) were purchased from ABclonal (Wuhan, China). Chemiluminescence substrate detection solution (#WBKLS0500) was purchased from MERCK (MilliPore, Boston, MA, USA). Cell Counting Kit-8 (CCK-8) (#BA00208) was obtained from Bioss (Beijing, China). Dimethylsulfoxide was purchased from MERCK (Sigma-Aldrich, St. Louis, MO, USA).

Yang S., Li S, & Chang J. (2022). Discovery of Cobimetinib as a novel A-FABP inhibitor using machine learning and molecular docking-based virtual screening. RSC Advances, 12(21), 13500-13510.

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Western Blot Analysis of Cell Signaling

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Protein samples were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to NC membranes, and blocked with TBST of 5% skim milk for 1 hour. The membranes were washed with TBST and then incubated with the specific primary antibody for 6 hours at 4°C. The membranes were then incubated for 1 hour at room temperature in secondary and the signal was detected by chemiluminescence (Bio rad, USA). The primary (1:1000) and secondary (1:10000) antibodies were purchased from Cell Signaling Technology, USA and listed as follows: GAPDH antibody(#2118), stat3 antibody(#12640), phospho-stat3 antibody(#98543), SAPK/JNK antibody(#9252), phospho-SAPK/JNK antibody(#4668), p65 antibody(#4764), phospho-p65 antibody(#3033), IKKα antibody(#2682), phospho-IKKα/β antibody(#2697), IκBα antibody(#4812), phospho-IκBα antibody(#2859), p38 MAPK antibody(#8690), phospho-p38 MAPK antibody(#4511), Erk1/2 antibody(#4695), phospho-Erk1/2 antibody(#4370) and HRP-linked goat anti-rabbit IgG(#7074).

Zhang C.X., Tu Y., Sun X.C., Chen D.G., Zhang W.N., Zhuang C.L., Wang Z.B, & Su L. (2022). Peramivir, an Anti-Influenza Virus Drug, Exhibits Potential Anti-Cytokine Storm Effects. Frontiers in Immunology, 13, 856327.

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Anti-inflammatory Mechanisms in Carrageenan-Induced Mice

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Male KM mice, weighing
16–18 g, were purchased from the Animal Laboratory Centre of
Jinzhou Medical University (Animal license number: SCXK [Liao] 2019-0007).
YXB (NO: 20210303) was supplied by China Resources in Liaoning Benxi
Third Pharmaceutical Co., Ltd. Celecoxib Capsules (Cele) (NO: DK1011)
were purchased from Pfizer. Carrageenan (NO: C804872) was purchased
from Shanghai Macklin Biochemical Co., Ltd. Isoflurane (NO: R510-22)
was purchased from RWD Life Science Co., Ltd. RIPA lysis buffer (NO:
P0013B), phosphatase inhibitor cocktail (NO: P1081), and phenylmethylsulfonyl
fluoride (PMSF) (NO: ST506) were purchased from Beyotime Biotechnology.
A Pierce Rapid Gold BCA Protein Assay Kit (NO: A53227) was purchased
from Thermo Scientific. SAPK/JNK Antibody (NO: 9252), Phospho-SAPK/JNK
Antibody (NO: 9251), and p38 MAPK Antibody (NO: 8690) were purchased
from Cell Signaling Technology. p-p38 Antibody (NO: WLP1576), ERK1/2
Antibody (NO: WL02371), p-ERK1/2 Antibody (NO: WLP1512), NF-κB
p65 Antibody (NO: WL01273b), p-NF-κB p65 Antibody (NO: WL02169),
and COX-2 Antibody (NO: WL01750) were purchased from Wanleibio Co.,
Ltd. β-actin Antibody (NO: AC026) and HRP Goat Anti-Rabbit IgG
(H + L) (NO: AS014) were purchased from ABclonal. An Ultra-High Sensitivity
ECL Kit (NO: BL523A) was purchased from Biosharp. Mouse TNF-α,
IL-1β, and IL-6 ELISA kits were purchased from R&D.

Hu X., Shao P., Liu X., Han L., Gui L., Cai Z., Qi M, & Dai C. (2022). Study on the Anti-Inflammatory Effect and Mechanism of Yuxuebi Tablet Based on Network Pharmacology. ACS Omega, 7(36), 32784-32794.

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