2 × 105 cells were seeded one day before the experiments and washed twice with PBS before they were lysed in RIPA buffer. Protein concentrations were determined by BCA assay. Protein samples were resolved for SDS-PAGE, blotted onto PVDF membranes and probed with antibodies for PTGS1 (Sigma, AV41836), GPS2 (custom-made rabbit polyclonal IgG, as previously described (36 (link),39 (link)), NCOR (Bethyl, A301-145A), SMRT (Bethyl, A301-147A), H3 (Abcam, ab1791), STAT6 (Sigma, S-6433), p-STAT6 (Thermo, 700247), β-actin (Abcam, ab8226), H3K27ac (Abcam, ab4729), H3K4me3 (Abcam, ab8580), H3K9me2 (Abcam, ab1220), H3K9me3 (Abcam, ab8898) or KDM1A (Abcam, ab17721). Membranes were developed using the ECL system. All antibodies are listed in the Supplementary Table II .
P stat6
Manufactured by Thermo Fisher Scientific
The P-STAT6 is a laboratory instrument designed for the detection and quantification of STAT6 protein phosphorylation. It utilizes an electrochemical detection method to provide precise and reliable measurements of STAT6 activation levels in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab1791, by Merck Group (1 mentions)
Ab8898, by Abcam (1 mentions)
Ab1220, by Abcam (1 mentions)
Ab8580, by Abcam (1 mentions)
Ab8226, by Abcam (1 mentions)
Ab4729, by Abcam (1 mentions)
Ab17721, by Abcam (1 mentions)
Annexin 5 apoptosis detection kit, by Thermo Fisher Scientific (1 mentions)
Perm buffer 3, by BD (1 mentions)
Phosflow, by BD (1 mentions)
2 protocols using p stat6
1
Protein Analysis of Cellular Signaling
2
Flow Cytometric Analysis of Neutrophil Markers
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Single‐cell suspensions of neutrophils were processed for analysis of extracellular and intracellular markers by flow cytometry, as previously described.13 , 23 Briefly, isolated neutrophils were stained for using fluorochrome‐conjugated mAbs directed against the following human antigens (from BioLegend unless otherwise stated; clone names in parentheses): CD11b (ICRF 44), CD16 (3G8), CD35 (E11), CD61 (VI‐PL2), CD63 (H5C6), CD66b (G10F5), CD62L (DREG‐56), CD66c (B6.2), CD114 (LMM741), CD124 (G077F6), CD132 (TUGh4), CD162 (KPL‐1), CD213a1 (SS12B), CD213a2 (SHM38), CXCR1 (8F1/CXCR1), CXCR2 (5E8/CXCR2), and CXCR4 (12G5). For intracellular staining, neutrophils were fixed with Fix Buffer I and permeabilized with Perm Buffer III (BD Phosflow, BD Bioscience), followed by intracellular staining for myeloperoxidase (MPO421‐8B2), phospho‐Y705 of STAT3 (pSTAT3; 13A3‐1), phospho‐Y694 of STAT5 (pSTAT5; SRBCZX, ThermoFisher), and phospho‐Y641 of STAT6 (pSTAT6; CHI2S4, ThermoFisher), as previously established.13 , 24 Cell viability was analyzed using Annexin V Apoptosis Detection Kit (eBioscience). Samples were acquired on a BD LSR Fortessa and analyzed using FlowJo software (Tristar).
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