Anti pjak1 tyr1034 1035

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-pJAK1 (Tyr1034/1035) is a primary antibody that detects phosphorylation of Janus kinase 1 (JAK1) at tyrosine residues 1034 and 1035. This antibody can be used to monitor JAK1 activation.

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3 protocols using anti pjak1 tyr1034 1035

Antibody-Based Cell Sorting Protocol

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Antibodies for cell sorting include anti-B220, anti-CD2, anti-CD3, anti-CD5, anti-CD8, anti-Gr-1, anti-CD41, anti-Ter119, anti-CD150, anti-CD48, anti-Kit and anti-Sca1 (Table S2). All antibodies were purchased from BioLegend. Anti-APC conjugated to paramagnetic microbeads were from Miltenyi Biotec. Antibodies for western blotting were anti-pStat5 (Y694), anti-Stat5 (4H1), anti-β-actin (8H10D10), anti-pJak2 (Tyr221), anti-Jak2, anti-pJAK1 (Tyr1034/1035) and anti-SOCS2 (Table S3), all purchased from Cell Signaling Technology. Anti-GAPDH was from Santa Cruz.

Jin X., Ng V., Zhao M., Liu L., Higashimoto T., Lee Z.H., Chung J., Chen V., Ney G., Kandarpa M., Talpaz M, & Li Q. (2022). Epigenetic downregulation of Socs2 contributes to mutant N-Ras-mediated hematopoietic dysregulation. Disease Models & Mechanisms, 15(5), dmm049088.

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Murine Cancer Cell Lines and Immunoblotting

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The mouse cancer cell lines B16 and LLC were purchased from ATCC (Manassas, VA). B16-OVA and MC38 cells were generously provided by Dr. Pan Zheng (University of Maryland; Baltimore, MD). All cell lines are mycoplasma free. The cells were maintained in Dulbecco’s modified Eagle’s medium or Roswell Park Memorial Institute 1640 medium (RPMI-1640) supplemented with 10% FBS, 1% penicillin-streptomycin, 2mM L-glutamine, and 1 mM sodium pyruvate at 37 °C in 5% CO₂.
For western blot, the anti-CNR2 antibody was purchased from Invitrogen (CA, USA). Anti-pJAK1-Tyr1034/1035, anti-pSTAT1-Ser727 antibodies, anti-STAT1, anti-pSTAT3-Ser727 antibodies, anti-STAT3, anti-Flag, and secondary antibodies HRP-mouse or HRP-rabbit were purchased from Cell Signaling Technology (Boston, Massachusetts USA). Anti-β-actin, anti-JAK1 were purchased from Proteintech (Chicago, IL, USA).
AEA and THC were purchased from Sigma (Sigma‐Aldrich, CA, USA). The anti-mouse PD-1 antibody was purified from hybridoma (clone G4; kindly provided by Dr. Lieping Chen, Yale University, New Haven, CT) culture supernatant.

Xiong X., Chen S., Shen J., You H., Yang H., Yan C., Fang Z., Zhang J., Cai X., Dong X., Kang T., Li W, & Zhou P. (2022). Cannabis suppresses antitumor immunity by inhibiting JAK/STAT signaling in T cells through CNR2. Signal Transduction and Targeted Therapy, 7, 99.

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Immunoblotting Analysis of Signaling Pathways

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The cells were lysed with NP-40 lysis buffer and centrifuged at 15,000 rpm for 15 min at 4 °C. The supernatants were collected, and total protein content was measured using the Bradford assay (Bio-Rad Laboratories). Lysates were separated with Bolt 4 to 12 % Bis-Tris polyacrylamide gels (Thermo Fisher Scientific), transferred to membrane filters, and subjected to immunoblotting using anti-GP130 (1:2000, Cell Signaling, #3732), anti-LIFR (1:1000, Santa Cruz Biotechnology, A-10), anti-EGFR (1:1000, Cell Signaling, #2232), anti-p-EGFR-Tyr1068 (1:1000, Cell Signaling, #2234), anti-AKT (1:1000, Cell Signaling, #4691), anti-p-AKT-Ser473 (1:1000, Cell Signaling, #4060), anti-ERK1/2 (1:1000, Cell Signaling, #9102), anti-p-ERK1/2-Thr202/Tyr204 (1:1000, Cell Signaling, #9101), anti-MEK1/2 (1:1000, Cell Signaling, #8727), anti-p-MEK1/2-Ser217/221 (1:1000, Cell Signaling, #9154), anti-Stat3 (1:1000, Cell Signaling, #4904), anti-p-Stat3-Tyr705 (1:1000, Cell Signaling, #9145), anti-Jak1 (1:1000, Cell Signaling, #3344), anti-p-Jak1-Tyr1034/1035 (1:1000, Cell Signaling, #74129), anti-Jak2 (1:1000, Cell Signaling, #3230), anti-p-Jak2-Tyr1008 (1:1000, Cell Signaling, #8082), anti-Jak3 (1:1000, Cell Signaling, #8827), anti-p-Jak3-Tyr980/981 (1:1000, Cell Signaling, #5031), and anti-beta-tubulin mouse antibody (1:5000, Sigma).

Hara T., Chanoch-Myers R., Mathewson N.D., Myskiw C., Atta L., Bussema L., Eichhorn S.W., Greenwald A.C., Kinker G.S., Rodman C., Castro L.N., Wakimoto H., Rozenblatt-Rosen O., Zhuang X., Fan J., Hunter T., Verma I.M., Wucherpfennig K.W., Regev A., Suvà M.L, & Tirosh I. (2021). Interactions between cancer cells and immune cells drive transitions to mesenchymal-like states in glioblastoma. Cancer cell, 39(6), 779-792.e11.

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