Stat1 d1k9y

Manufactured by Cell Signaling Technology

Sourced in United Kingdom

STAT1 (D1K9Y) is a primary antibody that recognizes the STAT1 protein. STAT1 is a transcription factor that mediates cellular responses to interferons, cytokines, and growth factors.

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Lab products found in correlation

Pstat1 tyr701, by Cell Signaling Technology (1 mentions) Ecl select reagent, by GE Healthcare (1 mentions) Alliance ld2, by Uvitec (1 mentions) Epcam, by Abcam (1 mentions) Bcl 2, by Abcam (1 mentions) Protein assay, by Bio-Rad (1 mentions) Irf1 d5e4, by Cell Signaling Technology (1 mentions) Polyvinylidene fluoride (pvdf), by Merck Group (1 mentions) Pstat1 y701 58d6, by Cell Signaling Technology (1 mentions) Gapdh hrp, by Cell Signaling Technology (1 mentions) Stat3 124h6, by Cell Signaling Technology (1 mentions) Pierce ripa buffer, by Thermo Fisher Scientific (1 mentions) Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Halt protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Ubiquitin, by Proteintech (1 mentions) Jak2 d2e12, by Cell Signaling Technology (1 mentions) Dnmt1, by Proteintech (1 mentions) Dnmt3a, by Proteintech (1 mentions) Aurora a 35c1, by Abcam (1 mentions) Jak1 6g4, by Cell Signaling Technology (1 mentions)

3 protocols using stat1 d1k9y

Western Blotting for Protein Analysis

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Western blotting was performed as previously reported [38 (link), 39 (link)]. Briefly, total protein extracts were obtained by lysing the cells in RIPA buffer (150 mM Sodium Chloride, 1% NP-40, 0.5% Sodium Deoxycholate, 0.1% SDS, and 50 mM TrisHCl pH 8.0) and proteins were quantified with the BCA method (Pierce, ThermoFisher Scientific); 20–50 μg of protein extracts were loaded on appropriate 7.5%, 10% or 12% polyacrylamide gels and SDS-PAGE was performed. Proteins were then transferred on Nitrocellulose membranes, which were probed over-night at 4 °C with specific antibodies diluted in 1% non-fat skim milk-PBS-T solution: ETV7/TEL2 (E-1, sc-374478), β-Tubulin (3F3-G2, sc-53140), STAT1 (D1K9Y, Cell Signaling Technology, Euroclone), p-STAT1 (Tyr701) (D4A7, Cell Signaling Technology), BCL-2 (100, sc-509), Survivin (D-8, sc-17779), EpCAM (ab71916, Abcam, Cambridge, UK), and HSP70 (C92F3A-5, sc-66048). Antibodies were obtained from Santa Cruz Biotechnologies (Heidelberg, Germany) when not explicitly indicated. Detection was performed with ECL Select reagent (GE Healthcare) using the UVITec Alliance LD2 (UVITec Cambridge, UK) imaging system.

Pezzè L., Meškytė E.M., Forcato M., Pontalti S., Badowska K.A., Rizzotto D., Skvortsova I.I., Bicciato S, & Ciribilli Y. (2021). ETV7 regulates breast cancer stem-like cell features by repressing IFN-response genes. Cell Death & Disease, 12(8), 742.

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Immunoblotting of Melanocyte Proteins

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Whole-cell extracts were lysed using Pierce RIPA buffer (Thermo Scientific) or NP-40 buffer supplemented with 1x Halt protease inhibitor cocktail (ThermoFisher) and 1x Halt phosphatase inhibitor cocktail (ThermoFisher). The concentration of cell extracts was measured by the Bio-Rad Protein Assay following manufacturer’s protocol. The same amounts of protein extracts were separated on the 4%−20% Mini-Protean TGX gel system (Bio-Rad) and transferred to PVDF (0.45 μm pore size, Millipore) membranes. Membranes were then incubated with the following primary antibodies: STAT1 (D1K9Y, 1:1000, Cell Signaling Technology), p-STAT1 (Y701, 58D6, 1:1000, Cell Signaling Technology), STAT3 (124H6, 1:1000, Cell Signaling Technology), p-STAT3 (Y705, D3A7, 1:2000, Cell Signaling Technology), IRF1 (D5E4, 1:1000, Cell Signaling Technology), TYR (PEP7h, 1:5000; gift from Vincent J. Hearing), TYRP1 (PEP1, 1:5000; gift from VJH), DCT (PEP8, 1:5000; gift from VJH), MITF (D5G7V, 1:1000, Cell Signaling Technology), PMEL/gp100 (EP4863(2), 1:1000, Abcam), and GAPDH-HRP (D16H11, 1:1000, Cell Signaling Technology). The secondary antibodies used for detection were HRP-conjugated goat anti-mouse and goat anti-rabbit IgG (1:5000, ThermoFisher). Band intensities of TIFF images were quantified by using Image J software.

Mo X., Raza Kazmi H., Preston-Alp S., Zhou B, & Raza Zaidi M. (2022). Interferon-gamma Induces Melanogenesis Via Post-Translational Regulation of Tyrosinase. Pigment cell & melanoma research, 35(3), 342-355.

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Alisertib Modulates Aurora A and Jak/STAT Signaling

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The effect of alisertib on Aurora A and the Jak/STATs signaling pathways was assessed by immunoblotting, as described previously 21 (link), 23 , using the following phospho-specific antibodies: p-Aurora A/B/C (Cell Signaling Technology, #2914), Aurora A (35C1) (Abcam, ab13824), p-Jak1 (T1022/1023) (Cell Signaling Technology, #3331), Jak1 (6G4) (Cell Signaling Technology, #3344), p-Jak2 (T1007/1008) (Cell Signaling Technology, #3776), Jak2 (D2E12) (Cell Signaling Technology, #3230), p-STAT1 (T701) (Cell Signaling Technology, #7649), STAT1 (D1K9Y) (Cell Signaling Technology, #14994), p-STAT2 (T690) (Cell Signaling Technology, #4441), STAT2 (D9J7L) (Cell Signaling Technology, #72604), PARP (46D11) (Cell Signaling Technology, #9532), DNMT3B (D7070) (Cell Signaling Technology, #67259), DNMT1 (Proteintech., 24206-1-AP), DNMT3A (Proteintech., 19366-1-AP), ubiquitin (Proteintech., 10201-1-AP), UHRF1(Proteintech., 21402-1-AP), flag (Abmart, M20008L) and β-actin (Proteintech.,66009-1-Ig).

Han J., Chen X., Xu J., Chu L., Li R., Sun N., Jiang Z., Liu H., Ge X., Zheng J., Yang J, & Ikezoe T. (2021). Simultaneous silencing Aurora-A and UHRF1 inhibits colorectal cancer cell growth through regulating expression of DNMT1 and STAT1. International Journal of Medical Sciences, 18(15), 3437-3451.

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