Phospho stat1 tyr701 58d6

Manufactured by Cell Signaling Technology

The Phospho-Stat1 (Tyr701) (58D6) antibody is a rabbit monoclonal antibody that recognizes Stat1 protein phosphorylated at tyrosine 701. It is designed for use in western blotting applications to detect the phosphorylation status of Stat1.

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Lab products found in correlation

Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Bca protein assay, by Thermo Fisher Scientific (2 mentions) Ucp1 ab10983, by Abcam (1 mentions) Fabp4 e71703 98, by Huabio (1 mentions) Cocktail 45 8099, by Thermo Fisher Scientific (1 mentions) Isg15, by Cell Signaling Technology (1 mentions) β actin hrp, by Cell Signaling Technology (1 mentions) Rps6 5g10, by Cell Signaling Technology (1 mentions) Pvdf membrane, by Thermo Fisher Scientific (1 mentions) Vybrant dyecycle ruby stain, by Thermo Fisher Scientific (1 mentions) Enzyme, by New England Biolabs (1 mentions) Stat2, by Santa Cruz Biotechnology (1 mentions) Stat5b, by Santa Cruz Biotechnology (1 mentions) Tofacitinib citrate, by Merck Group (1 mentions) Rabbit igg, by EpiCypher (1 mentions) Phospho stat1 ser727, by Cell Signaling Technology (1 mentions) Stat1, by Cell Signaling Technology (1 mentions) Stat3, by Santa Cruz Biotechnology (1 mentions) Ifn γ, by Merck Group (1 mentions) Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions)

4 protocols using phospho stat1 tyr701 58d6

Protein Isolation and Western Blot Analysis

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Total protein was isolated from cells or tissues using RIPA buffer supplemented with protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA, USA). The protein concentration was determined with the BCA protein assay (Thermo Fisher Scientific, USA). Protein separation and western blot analysis were conducted as described earlier [5 (link)]. GADD45A antibody (sc-6850, 1: 1,000) was from Santa Cruz Biotechnology (Santa Cruz). Perilipin-1 (ab61682, 1:2000) and UCP1 (ab10983, 1:1000) were from Abcam. FABP4 (E71703-98, 1:2000) and GAPDH (EM1101, 1:5000) were from HuaBio. Cocktail (45-8099, 1:2000) was from Thermo Fisher Scientific. Stat1 (D4Y6Z, 1: 1000) and Phospho-Stat1 (Tyr701) (58D6, 1:1000) were obtained from Cell Signaling Technology. The horseradish peroxidase (HRP)-conjugated secondary antibody (anti-rabbit IgG, 111-035-003 or anti-mouse IgG; 115-035-003, Jackson ImmunoResearch) was diluted 1:10,000. Immunodetection was performed using enhanced chemiluminescence western blotting substrate (Google Biotechnology, Wuhan, Hubei, China) and detected by ChemiScope3500 Mini System.

You W., Liu S., Li J., Tu Y, & Shan T. (2023). GADD45A regulates subcutaneous fat deposition and lipid metabolism by interacting with Stat1. BMC Biology, 21, 212.

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Immunoblotting of Innate Immune Signaling

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Cells were lysed in RIPA buffer (+ 1x HALT protease and phosphatase
inhibitor and 10–30 μg total protein from whole cell lysates was
run on SDS-PAGE and transferred to PVDF membranes (Thermo Scientific). The
membranes were probed in 5 % BSA or milk in PBS-T (Phosphate-buffered
saline/Tween 20) for ZAP (N3C2, GeneTex), ISG15 (Cell Signaling, #2743), OAS1
(D1W3A, Cell Signaling), RIG-I (Alme-1, AdipoGen), IRF1 (D5E4, Cell Signaling),
IRF3 (D83B9, Cell Signaling), phospho-IRF3 (Ser386) (EPR2346, Abcam), STAT1
(42H3, Cell Signaling), phospho-STAT1 (Tyr701) (58D6, Cell Signaling), RPS6
(5G10, Cell Signaling), CSTF2 (Bethyl Laboratories), Calnexin (C5C9, Cell
Signaling), FLAG (M2, Sigma), Myc (71D10, Cell Siganling) or β-Actin-HRP
(13E5, Cell Signaling). For detailed information about the source of the
antibodies and dilutions used please refer to the Life Sciences Reporting
Summary.

Schwerk J., Soveg F.W., Ryan A.P., Thomas K.R., Hatfield L.D., Ozarkar S., Forero A., Kell A.M., Roby J.A., So L., Hyde J.L., Gale M J.r., Daugherty M.D, & Savan R. (2019). RNA-binding protein isoforms ZAP-S and ZAP-L have distinct antiviral and immune resolution functions. Nature immunology, 20(12), 1610-1620.

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Multiplexed Signaling Pathway Analysis

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All chemicals, unless otherwise noted, were obtained from ThermoFisher or Merck. Enzymes were obtained from New England Biolabs. The following drugs/dyes were used for this work: IFNγ (final concentration of 50 ng/ml, Merck), dTAG13 (final concentration of 100 nM, Merck), Vybrant DyeCycle Ruby Stain (final concentration of 5 μM, ThermoFisher) and Tofacitinib citrate also known as CP‐690550 (JAK inhibitor, concentration of 10 μM, Merck). The following antibodies were used for this work: GBP5 (Cell Signaling Technology, 67798; Abcam, ab96119), STAT1 (Cell Signaling Technology, 9172), Phospho‐STAT1 (Ser727) (Cell Signaling Technology, 9177), Phospho‐STAT1 (Tyr701) (58D6) (Cell Signaling Technology, 9167), STAT2 (Santa Cruz Biotechnology, sc‐1668), STAT3 (Santa Cruz Biotechnology, sc‐8019), STAT5B (Santa Cruz Biotechnology, sc‐1656), IRF1 (Cell Signaling Technology, 8478), IRF9 (ThermoFisher Scientific, 702322), a‐TUB (Merck, T9026), anti‐rabbit (fluorophore‐conjugated) (LI‐COR, 926‐32211), anti‐mouse (fluorophore‐conjugated) (Rockland, 610‐744‐124), Rabbit IgG (Epicypher, 13‐0042k).

Tehrani S.S., Mikulski P., Abdul‐Zani I., Mata J.F., Siwek W, & Jansen L.E. (2023). STAT1 is required to establish but not maintain interferon‐γ‐induced transcriptional memory. The EMBO Journal, 42(14), e112259.

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Western Blot Analysis of DNA Damage Signaling

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Cells were detached with Accutase (Sigma), collected and lysed in RIPA buffer (Pierce) supplemented with complete phosphatase and protease inhibitors (Cell signaling). Proteins were quantified by BCA (ThermoFisher Scientific) and 20 µg of proteins were separated by gel electrophoresis in NuPAGE 4–12% Bis-Tris Gel Novex (ThermoFisher Scientific) with NuPAGE 1x MOPS buffer (Life technologies). Proteins were transferred to nitrocellulose membrane (Life technologies) by iBlot^TM 2 Transfer Device (ThermoFisher Scientific). Membranes were blocked with Odyssey blocking buffer PBS (LI-COR) and probed with primary antibodies as indicated for 16 hours at 4°C at a concentration of 1:500 for Phospho-histone H2A.X (Ser139) (Cell Signaling 2577), Phospho-P-53 (Ser 15) (Cell Signaling 9284), Phospho-STAT1 (Tyr701) (58D6) (Cell Signaling 9167), STING (D2P2F) (Cell Signaling 13647) and cGAS (D1D3G) (Cell Signaling 15102), P-S1981-ATM [EP1890Y] (ABCAm 81292) and GAPDH (Millipore MAB374), followed by either IRDye680LT goat anti-rabbit (LI-COR, 926–68021) or IRDye800CW goat anti-mouse (LI-COR, 925–32210) as per the manufacturer’s instructions and analyzed in LI-COR Odyssey.

Lopez-Pelaez M., Young L., Vazquez-Chantada M., Nelson N., Durant S., Wilkinson R.W., Poon E., Gaspar M., Valge-Archer V., Smith P, & Dovedi S.J. (2022). Targeting DNA damage response components induces enhanced STING-dependent type-I IFN response in ATM deficient cancer cells and drives dendritic cell activation. Oncoimmunology, 11(1), 2117321.

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