Cells were stimulated for 25 min in the presence or absence of 25 ng/ml recombinant human IL-12 (R&D Systems), 50 ng/ml IL-23 (eBioscience) or 25 ng/ml IL-1β (R&D Systems). Cells were then washed two times in FACS buffer and fixed for 20 min in BD Fix and Perm buffer at room temperature (BD Biosciences), washed in FACS buffer, and fixed in 90% ice-cold methanol for 30 min on ice. Cells were washed two times with BD Perm/Wash and stained with antibodies against intracellular markers, including pSTAT4 (Y693) and pSTAT3 (pY705) (both from BD Biosciences) or Ikbα (Cell Signaling), in BD Perm/Wash for 40 min at room temperature. Cells were then washed two times, resuspended in BD Perm/Wash and analyzed by flow cytometry.
Pstat3 py705
Manufactured by BD
Sourced in United States
PSTAT3 (pY705) is a primary antibody that detects phosphorylated STAT3 at tyrosine 705. It is used for the detection and quantification of phosphorylated STAT3 in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Perm wash, by BD (1 mentions)
Recombinant human il 12, by R&D Systems (1 mentions)
Il 23, by Thermo Fisher Scientific (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Golgiplug, by BD (1 mentions)
Annexin 5 apoptosis detection kit, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Vcam 1, by Thermo Fisher Scientific (1 mentions)
Il 17, by Thermo Fisher Scientific (1 mentions)
Icam 1, by Thermo Fisher Scientific (1 mentions)
Golgistop, by BD (1 mentions)
Il 27, by Thermo Fisher Scientific (1 mentions)
P stat1 py701, by BD (1 mentions)
Cd106, by BD (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by BD (1 mentions)
Il 17, by BD (1 mentions)
Cd11a, by BD (1 mentions)
2 protocols using pstat3 py705
1
Cytokine-Induced Signaling Pathway Analysis
2
Investigating Immune Cell Signaling Pathways
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The following antibodies were used as follow: For flow cytometry determination, CD3, CD8, CD54 (intercellular adhesion molecule [ICAM]-1), CD106 (vascular adhesion molecule [VCAM]-1), CD11a (lymphocyte function-associated antigen-1 [LFA-1]), CD29 (integrin β1), IL-27, IFN-γ, IL-17, phosphorylated (p)-STAT1 (pY701), p-STAT1 (pS727), p-STAT2 (pY689), p-STAT3 (pS727), p-STAT3 (pY705), p-STAT4 (pY693), p-STAT5 (pY694), p-STAT6 (pY641), TCCR-WSX-1 (IL-27 receptor), IL-17 receptor (R), IFN-γR1, and Ki-67 mAbs were purchased from BD Biosciences (Franklin Lakes, USA), eBioscience (San Diego, USA), or R and D systems (Minneapolis, USA); for cytokine neutralization, IL-27, IFN-γ, IL-17, ICAM-1 and VCAM-1 mAbs were from eBioscience.
Recombinant human IL-27, IFN-γ, IL-17A were purchased from R and D systems or eBioscience. STAT1 inhibitor S14-95 and STAT3 inhibitor Galiellalactone were from Enzo Life Sciences (Farmingdale, USA). Annexin V Apoptosis Detection Kit was from eBioscience. PMA and ionomycin were from Sigma-Aldrich (St. Louis, USA), and GolgiPlug and GolgiStop were from BD Bioscience. Foxp3 fixation/permeabilization concentrate and diluent were from eBioscience. Fluorescent dye CFSE was from Invitrogen (Carlsbad, USA).
Recombinant human IL-27, IFN-γ, IL-17A were purchased from R and D systems or eBioscience. STAT1 inhibitor S14-95 and STAT3 inhibitor Galiellalactone were from Enzo Life Sciences (Farmingdale, USA). Annexin V Apoptosis Detection Kit was from eBioscience. PMA and ionomycin were from Sigma-Aldrich (St. Louis, USA), and GolgiPlug and GolgiStop were from BD Bioscience. Foxp3 fixation/permeabilization concentrate and diluent were from eBioscience. Fluorescent dye CFSE was from Invitrogen (Carlsbad, USA).
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