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Anti pstat3 tyr705

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Anti‐pSTAT3 (Tyr705) is a primary antibody that recognizes the phosphorylated form of STAT3 at tyrosine 705. STAT3 is a transcription factor that plays a key role in cellular signaling pathways.

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Lab products found in correlation

Anti stat3 antibody, by Abcam (2 mentions) Protease phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Anti p stat5, by Cell Signaling Technology (1 mentions) Halt protease phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Anti stat5 antibody, by Cell Signaling Technology (1 mentions) Ripa lysis buffer, by Avantor (1 mentions) Anti stat3, by Cell Signaling Technology (1 mentions) Dako autostainer plus en vision kit, by Agilent Technologies (1 mentions) Rabbit anti mmp 9, by Abcam (1 mentions) Prim 1640 medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions) Mouse anti bcl 2 c 2, by Santa Cruz Biotechnology (1 mentions) Mouse anti β actin, by OriGene (1 mentions) Rabbit anti mmp 2, by Affinity Biosciences (1 mentions) Fetal bovine serum (fbs), by Lonsera (1 mentions) Anti jak2, by Abcam (1 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Anti stat3, by Abcam (1 mentions)

5 protocols using anti pstat3 tyr705

1

Leptin-Induced STAT3/STAT5 Phosphorylation

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LepR-Ba/F3 cells were incubated in serum-free medium overnight and then treated with 100 ng/mL leptin (#Z02962-1; GenScript, Nanjing, China) or 500 ng/mL agonist-type antibody at 37 °C, 5% CO2 for 30 min. The cells were collected and washed once with ice-cold PBS containing a Halt protease/phosphatase inhibitor cocktail (#78446; Thermo Scientific, Waltham, MA, USA), followed by 30 min of lysis with RIPA buffer containing a protease/phosphatase inhibitor cocktail on ice with shaking. The resultant supernatant was collected by centrifuging at 14,000× g rpm for 10 min. The amount of total STAT3 or STAT5 and phosphorylated STAT3 or STAT5 were determined by a western blot analysis of the cell lysates’ supernatant using anti-STAT3 antibody (#32500; Abcam, Cambridge, UK), anti-STAT5 antibody (#9363T; Cell Signaling Technology), anti-pSTAT3 (Tyr705) (#76315; Abcam), and anti-pSTAT5 (#4322P; Cell Signaling Technology, Danvers, MA, USA), respectively.
Wang T., Chen X., Yang G, & Shi X. (2024). Selection of Leptin Surrogates by a General Phenotypic Screening Method for Receptor Agonists. Biomolecules, 14(4), 457.
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2

Leptin-Induced STAT3 Activation Assay

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Cells were cultured in a serum‐free medium overnight, then treated with serum‐free medium containing 10 × 10−9m leptin, 10 × 10−9m agonist antibody at 37 °C, 5% CO2 for 30 min. Cells were next collected and washed once with ice‐cold PBS containing Halt protease and a phosphatase inhibitor mixture (Pierce). The resulting cells were lysed on ice for 30 min with a RIPA lysis buffer (Amresco) containing Halt protease and phosphatase inhibitor mixture. Cell debris was removed by centrifugation at 14 000 × g for 10 min. The amount of total STAT3 and phosphorylated STAT3 was determined by western‐blot analysis of the cell lysates supernatant using anti‐STAT3 antibody (abcam) and anti‐pSTAT3 (Tyr705) (abcam), respectively.
Tao P., Kuang Y., Li Y., Li W., Gao Z., Liu L., Qiang M., Zha Z., Fan K., Ma P., Friedman J.M., Yang G, & Lerner R.A. (2020). Selection of a Full Agonist Combinatorial Antibody that Rescues Leptin Deficiency In Vivo. Advanced Science, 7(16), 2000818.
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3

Prostate Cancer Cell Lysate Analysis

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Prostate cancer cell lysates were prepared and analyzed by Western blot analysis as in a previous publication.22 Primary antibodies used were anti‐STAT3 (#4904; Cell Signaling Technology), anti‐pSTAT3 Tyr‐705 (ab76315; Abcam) and anti‐beta‐actin (#691001; BioMedicals MP) was used as loading control.
Hellsten R., Lilljebjörn L., Johansson M., Leandersson K, & Bjartell A. (2019). The STAT3 inhibitor galiellalactone inhibits the generation of MDSC‐like monocytes by prostate cancer cells and decreases immunosuppressive and tumorigenic factors. The Prostate, 79(14), 1611-1621.
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4

Immunostaining of STAT3 Activation

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Suspensions of monocytes cultured for 72 hours in CM from prostate cancer cells or MDSCs isolated from mCRPC patients were placed in a cytospin funnel clamped to a glass slide and spun at 800 rpm for 2 minutes. The slides were air‐dried, fixed in 4% formaldehyde for 10 minutes, and permeabilized with 1% Triton‐X Tris buffer, pH 7.6 for 1 hour. The slides were subjected to immunohistochemistry using Dako Autostainer Plus En VisionTM+ Kit (Dako). Cells were stained using the antibody anti‐pSTAT3 Tyr‐705 (ab76315; Abcam).
Hellsten R., Lilljebjörn L., Johansson M., Leandersson K, & Bjartell A. (2019). The STAT3 inhibitor galiellalactone inhibits the generation of MDSC‐like monocytes by prostate cancer cells and decreases immunosuppressive and tumorigenic factors. The Prostate, 79(14), 1611-1621.
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5

Salidroside Modulates Apoptosis and Signaling Pathways

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PRIM1640 medium, penicillin‐streptomycin solution and trypsin‐EDTA were purchased from Gibco (California, USA). Fetal bovine serum (FBS) was purchased from Lonsera (Montevideo, Uruguay). Salidroside (pure ≥98%) was purchased from Tauto Biotech Co., Ltd. (Shanghai, China) and its product ID is E‐0069. Salidroside was dissolved in phosphate buffer solution (PBS) and filtered through a 0.22‐μm filter before use. Annexin V apoptosis detection kit was purchased from NeoBioscience (Shenzhen, China). Antibodies were obtained from the following sources: rabbit anti‐MMP9 (#ab38898), anti‐pSTAT3 (Tyr705) (#ab76315), anti‐STAT3 (#ab68153), anti‐JAK2 (#ab108596) and anti‐pJAK2 (Y1007 + Y1008) (#ab32101) from Abcam, rabbit anti‐MMP2 (#AF0577) from Affinity, mouse anti‐Bcl‐2 (C‐2) (#sc‐7382) from Santa Cruz Biotechnology, rabbit anti‐Bax (#GB11007) from Servicebio, rabbit anti‐cleaved caspase 3 (#9664) from Cell Signaling Technology, mouse anti‐β‐Actin (#LocusID60) from OriGene, rabbit anti‐GAPDH (#10494‐1‐AP) and HRP‐conjugated secondary anti‐rabbit IgG antibody (#SA00001‐2) from Proteintech.
Shang H., Wang S., Yao J., Guo C., Dong J, & Liao L. (2019). Salidroside inhibits migration and invasion of poorly differentiated thyroid cancer cells. Thoracic Cancer, 10(6), 1469-1478.
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