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Phospho jak2

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Phospho-JAK2 is a lab equipment product from Cell Signaling Technology. It is designed to detect the phosphorylation of the JAK2 protein, which plays a crucial role in cellular signaling pathways.

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Lab products found in correlation

Phospho stat3, by Cell Signaling Technology (26 mentions) Stat3, by Cell Signaling Technology (13 mentions) Phospho stat1, by Cell Signaling Technology (10 mentions) Phospho jak1, by Cell Signaling Technology (9 mentions) Stat1, by Cell Signaling Technology (8 mentions) Phospho akt, by Cell Signaling Technology (8 mentions) Phospho stat5, by Cell Signaling Technology (7 mentions) Phospho stat2, by Cell Signaling Technology (7 mentions) Phospho tyk2, by Cell Signaling Technology (6 mentions) Phospho p38, by Cell Signaling Technology (6 mentions) Phospho jnk, by Cell Signaling Technology (6 mentions) Gapdh, by Cell Signaling Technology (5 mentions) Bcl xl, by Cell Signaling Technology (5 mentions) Phospho erk1 2, by Cell Signaling Technology (5 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (4 mentions) Caspase 3, by Cell Signaling Technology (4 mentions) Caspase 9, by Cell Signaling Technology (4 mentions) Cleaved caspase 8, by Cell Signaling Technology (4 mentions) Phospho src, by Cell Signaling Technology (4 mentions) Phospho erk, by Cell Signaling Technology (4 mentions)

Spelling variants of this product

Anti-phospho-JAK2 (27 citations) Phospho-JAK2 (Tyr1007/1008), (10 citations) Anti-phospho-JAK2 (Tyr1007/1008), (7 citations) Anti–phospho-JAK2 (Y1007/1008) (3 citations) Phospho-JAK2 antibody (2 citations) Anti-phospho-JAK2 (Tyr1007/Tyr1008), (2 citations) Phospho-JAK2 mAb (2 citations) Phospho‐ Y1007/1008‐JAK2 (2 citations) Rabbit anti-phospho-JAK2 (2 citations)

45 protocols using phospho jak2

1

Western Blot Analysis of Apoptosis Signaling

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CCA cell lines were lysed by RIPA lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 0.1% SDS, 1% TritonX-100, 1% sodium deoxycholate). Protein samples (20 µg/lane) were subjected to electrophoresis in 12% sodium dodecyl sulfate‑polyacrylamide gel (SDS-PAGE) and transferred onto nitrocellulose membranes. After blocking with 5% skimmed milk at room temperature for 1 h, blots were incubated with respective primary antibodies at 4 °C overnight and subsequently horseradish peroxidase (HRP)‑conjugated secondary antibody at room temperature for 1 h (1:10,000). Antibody complexes were visualized using an enhanced chemiluminescence detection system (ECL kit; 1705061) from Bio-Rad Laboratories. Primary antibodies were purchased from Cell Signaling Technology: Bcl-2-associated X protein; Bax (5023T), b-cell lymphoma protein 2; Bcl-2 (4223T), cleaved caspase-3 (9664T), cleaved poly (ADP-ribose) polymerase; PARP (5625T), signal transducer and activator of transcription 3; STAT3 (12640S), phospho-STAT3 (9145T), Janus kinase; JAK2 (3230T) and phospho-JAK2 (3776S). Beta-actin (β-actin) from Proteintech (catalog no. 66009-1-Ig) was used as a loading control.
Jantalika T., Manochantr S., Kheolamai P., Tantikanlayaporn D., Saijuntha W., Pinlaor S., Chairoungdua A., Paraoan L, & Tantrawatpan C. (2022). Human chorion-derived mesenchymal stem cells suppress JAK2/STAT3 signaling and induce apoptosis of cholangiocarcinoma cell lines. Scientific Reports, 12, 11341.
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2

EGCG Modulation of Inflammatory Pathways

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EGCG (purity > 95%) was obtained from Huzhou Rongkai Foliage Extract Co., Ltd. (Huzhou, China). A fatty acid-albumin complex solution containing palmitic acid (PA, Sigma–Aldrich, St Louis, MO, USA) and fatty acid-free bovine serum albumin (BSA, Sigma, St Louis, MO, USA) was prepared as described previously [44 (link)]. All cell culture reagents were purchased from Gibco (Grand Island, NY, USA). Primary antibodies specific for NF-κB, phospho-NF-κB, IκB-α, phospho-IκB-α, Stat3, phosphoStat3, Jak2, and phospho-Jak2 were purchased from Cell Signaling Technology (Beverly, MA, USA), and Iba-1 was obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). D12450J normal chow diet and D12492 high-fat diet were purchased from Research Diets, Inc. Co., Ltd. (New Brunswick, NJ, USA).
Mao L., Hochstetter D., Yao L., Zhao Y., Zhou J., Wang Y, & Xu P. (2019). Green Tea Polyphenol (−)-Epigallocatechin Gallate (EGCG) Attenuates Neuroinflammation in Palmitic Acid-Stimulated BV-2 Microglia and High-Fat Diet-Induced Obese Mice. International Journal of Molecular Sciences, 20(20), 5081.
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3

Curcumin and Cisplatin Synergistic Pathway

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Curcumin, cisplatin, IL6, and all other chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA). Phospho-STAT3, STAT3, phospho-JAK2, JAK2, phospho-H2AX, phospho-CXCR4, Caspase-9, Cleaved Caspase-9, Cleaved caspase-8, Caspase-3, Cleaved Caspase-3, PARP, XIAP, cIAP1, cIAP2, Bcl-xL, Bax, p27, p21, Survivin, c-Myc, Nanog, Aldh, Gapdh, etc., were purchased from Cell Signaling Technologies (3 Trask Lane, Danvers, MA, USA). SOX2, Hsp60, and Bcl2 antibodies were procured from Santa Cruz Biotechnology (Finnell Street Dallas, TX, USA). Annexin V-FITC, propidium iodide staining solution, Hoechst 33342 Solution, BD Cytofix/Cytoperm plus fixation, and apermeabilization solution kit (a BD MitoScreen (JC-1) Kit) were purchased from BD Biosciences (Qume Drive, San Jose, CA, USA).
Khan A.Q., Ahmed E.I., Elareer N., Fathima H., Prabhu K.S., Siveen K.S., Kulinski M., Azizi F., Dermime S., Ahmad A., Steinhoff M, & Uddin S. (2020). Curcumin-Mediated Apoptotic Cell Death in Papillary Thyroid Cancer and Cancer Stem-Like Cells through Targeting of the JAK/STAT3 Signaling Pathway. International Journal of Molecular Sciences, 21(2), 438.
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4

Viscum album Extract Modulates STAT3 Pathway

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We obtained an extract of Viscum album, a parasitic plant that grows on Malus domestica (referred to as VaM) from Dalim BioTech (Wonju, Republic of Korea), which produces it as a product called Helixor M. Doxorubicin was purchased from Selleck Chem (Munich, Germany). Primary antibodies against STAT3, Phospho-STAT3 (Tyr705, Cat No. 9145s), Src, Phospho-Src, Phospho-Jak2, cleaved-caspase 3, and cleaved-PARP were purchased from Cell Signaling Technology (Beverly, MA, USA). Jak2, β-actin (Cat No. sc-47778), and secondary antibodies (Cat No. sc-516102 and sc-2004) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).
Park Y.R., Jee W., Park S.M., Kim S.W., Bae H., Jung J.H., Kim H., Kim S., Chung J.S, & Jang H.J. (2023). Viscum album Induces Apoptosis by Regulating STAT3 Signaling Pathway in Breast Cancer Cells. International Journal of Molecular Sciences, 24(15), 11988.
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5

Characterizing IL23R Signaling Pathways

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Proteins were detected by flow cytometry with Alexa Fluor 488 or phycoerythrin-labelled antibodies to IL23R (FAB41001P, R&D Systems), phospho-JAK1, phospho-JAK2, phospho-JAK3, phospho-TYK2, phospho-STAT1, phospho-STAT2, phospho-STAT3, phospho-STAT5, phospho-STAT6, FLAG (Cell Signaling) or phospho-STAT4 (BD Biosciences).
IL23R was immunoprecipitated using anti-IL23R antibody (Santa Cruz Technology, Santa Cruz, CA) bound to Protein A or Protein G Sepharose (EMD Millipore, Billerica, MA). Associated proteins were blotted with antibodies to JAK2, STAT3 (Cell Signaling), TYK2 (Abcam, Cambridge, MA), IL12Rβ1 (EMD Millipore), JAK3 (Santa Cruz Biotechnology) or IL23A (Proteintech). Control proteins were examined with glyceraldehyde 3-phosphate dehydrogenase or IL23R antibodies (EMD Millipore) as per ref 41 (link).
Sun R., Hedl M, & Abraham C. (2019). IL23 induces IL23R recycling and amplifies innate receptor-induced signalling and cytokines in human macrophages, and the IBD-protective IL23R R381Q variant modulates these outcomes. Gut, 69(2), 264-273.
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6

NLRP3 Inflammasome Activation in Macrophages

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All culture materials were purchased from Gibco (Grand Island, NY). LY294002 (PI3K/Akt inhibitor) and rapamycin (p70S6K inhibitor) were purchased from Calbiochem (La Jolla, CA). Mouse monoclonal antibodies (mAB) against SREBP-1c, NLRP3, IL-1β, pro-IL-1β, caspase-1, pro-caspase-1, phospho-Akt, and Akt were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit polyclonal antibodies against phospho-p70S6K, p70S6K, phospho-JAK2, and JAK2 were purchased from Cell Signaling Technology (Beverly, MA). SREBP-1c, NLRP3, JAK2 siRNA and control siRNA were purchased from the National RNAi Core Facility in Academic Sinica, Taipei, Taiwan. The integrin β1-siRNA were purchased from Invitrogen (Carlsbad, CA). P. gingivalis lipopolysaccharide (Pg-LPS) was purchased from InvivoGen (San Diego, CA). Other chemicals of reagent grade were obtained from Sigma (St. Louis, MO).
Kuo H.C., Chang L.C., Chen T.C., Lee K.C., Lee K.F., Chen C.N, & Yu H.R. (2016). Sterol Regulatory Element-Binding Protein-1c Regulates Inflammasome Activation in Gingival Fibroblasts Infected with High-Glucose-Treated Porphyromonas gingivalis. Frontiers in Cellular and Infection Microbiology, 6, 195.
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7

Antibody-Based Protein Expression Analysis

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Antibodies: Phospho-Smad2 (#3108, Cell Signaling Tech. 1/1000), phospho-Smad3(#9520, Cell Signaling Tech. 1/1000), phospho-Stat3 (#9131, Cell Signaling Tech. 1/1000), phospho-Jak2 (#3771, Cell Signaling Tech. 1/1000), Snail (#3879, Cell Signaling Tech. 1/1000), EEA1 (#C45B10, Cell Signaling Tech. 1/100) and total Stat3(#4904, Cell Signaling Tech. 1/1000) were purchased from Cell Signaling Technology; SMAD2/3 (610843, BD Biosciences, 1/1000), anti-myc (sc789, 1/1000 and 9E10, sc40, 1/1000), TβRI (sc398, 1/500) Lamin B (sc6217, 1/1000) TβRII (sc17792, 1/500), FKBP12 (sc6174, 1/500), GAPDH (sc32233, 1/1000) and GPR50 (sc50590, 1/500) came from Santa Cruz Biotechnology. Monoclonal and polyclonal Flag-Antibodies were used from Sigma-Aldrich (M2-F3165, 1/1000, and F7425, 1/1000). HA (11867423001, 1/1000) and GFP (11814460001, 1/2000) antibodies were used from Roche. Anti-Tubulin was purchased from AbD Serotec (MCA77G, 1/2000). GPR50 antibody7 was produced by Kernov Antibody Services (1/1000)49 (link). Polyclonal Anti GPR50 (H00009248-B01P, 1/500) was purchased from Novus Biologicals. Anti TGFβRI-pS165 (ab112095, 1/500) was purchased from Abcam. All antibodies were employed according to the recommended dilutions for either immunoprecipitation or western blotting.
Reagents: FK506 and SB431542 were purchased from Sigma Aldrich and recombinant TGFβ-1 was purchased from Peprotech.
Wojciech S., Ahmad R., Belaid-Choucair Z., Journé A.S., Gallet S., Dam J., Daulat A., Ndiaye-Lobry D., Lahuna O., Karamitri A., Guillaume J.L., Do Cruzeiro M., Guillonneau F., Saade A., Clément N., Courivaud T., Kaabi N., Tadagaki K., Delagrange P., Prévot V., Hermine O., Prunier C, & Jockers R. (2018). The orphan GPR50 receptor promotes constitutive TGFβ receptor signaling and protects against cancer development. Nature Communications, 9, 1216.
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8

Western Blot and qRT-PCR Analysis of Signaling Pathways

Cited 2 times
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The protein samples from stimulated or transfected cells were extracted by using the Laemmli Sample Buffer (Bio-Rad Laboratories, Inc., USA). The concentrations of protein samples were quantified by using the BCA™ Protein Assay Kit (Beyotime #P0012). Antibodies purchased from Cell Signaling Technology (Beverly, MA) included: phospho-STAT3 (Tyr705) (#9145), phospho-STAT3 (Ser727) (#49081), STAT3 (#4904), phospho-JAK2 (Tyr1007/1008) (#3771), JAK2 (#3230), phospho-JAK1 (Tyr1034/1035) (#74129), JAK1 (#3344), phospho-TYK2 (Tyr1054/1055) (#68790), TYK2 (#14193), phospho-STAT1 (Tyr701) (#7649), STAT1 (#14994), phospho-AKT (Ser473) (#4060), AKT(Pan) (#4821), phospho-NF-κB p65 (Ser536) (#3033), NF-κB p65 (#8242), cyclin D1 (#2978), cyclin A2 (#4656), cyclin B1 (#12231), Caspase-3 (#29629), Bcl-xL (#2762), and Survivin (#2803). PARP antibody and GAPDH antibody were purchased from Beyotime (#AP102) and Genscript (#A00192), respectively. WB analysis was conducted as previously described58 (link). Three independent experiments were performed using samples collected at different days. qRT-PCR was conducted as described previously59 (link). All the primers used were listed in Fig. S1b.
She S., Zhao Y., Kang B., Chen C., Chen X., Zhang X., Chen W., Dan S., Wang H., Wang Y.J, & Zhao J. (2020). Combined inhibition of JAK1/2 and DNMT1 by newly identified small-molecule compounds synergistically suppresses the survival and proliferation of cervical cancer cells. Cell Death & Disease, 11(9), 724.
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9

Signaling Pathway Protein Analysis

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Anti β-Actin was obtained from Abcam and used at a dilution of 1:2000. Anti phospho-JAK1 (Tyr 1022/1023) was obtained from Millipore and used at a dilution of 1:1000. All other antibodies—JAK1, JAK2, STAT1, STAT3, STAT5, phospho-JAK2 (Tyr 1007/1008), phospho-STAT1 (Tyr 701), phospho-STAT3 (Tyr 705), phospho-STAT5 (Tyr 694), phospho-c-Jun (Ser 73), phospho-ERK1/2 (Erk1 Thr 202 and Tyr 204, Erk2 Thr 185 and Tyr 187) and phospho-Akt (Thr 308)—were obtained from Cell Signalling Technology and used at a dilution of 1:1000.
Thomas S., Fisher K.H., Snowden J.A., Danson S.J., Brown S, & Zeidler M.P. (2015). Methotrexate Is a JAK/STAT Pathway Inhibitor. PLoS ONE, 10(7), e0130078.
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10

Intracellular Phospho-Protein Analysis

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Intracellular proteins were detected in permeabilized cells by flow cytometry with Alexa Fluor 647-, phycoerythrin- or Alexa Fluor 488-labeled antibodies to phospho-JAK1, phospho-JAK2, phospho-JAK3, phospho-TYK2, phospho-ERK, phospho-p38, phospho-JNK and phospho-IκBα (Cell Signaling Technology, Danvers, MA).
Hedl M., Proctor D.D, & Abraham C. (2016). JAK2 Disease-Risk Variants are Gain of Function and JAK Signaling Threshold Determines Innate Receptor-Induced Proinflammatory Cytokine Secretion in Macrophages. Journal of immunology (Baltimore, Md. : 1950), 197(9), 3695-3704.
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