Mouse anti igg1 pe antibody

For Ki67-based cell cycle analysis, BMMNCs were surface stained to distinguish HSPC fractions. After fixation and permeabilization using Cytofix/Cytoperm Kit (BD Bioscience), according to the manufacturer's protocol, cells were stained with an anti-Ki67 antibody (BD Bioscience) and Hoechst 33342 (Invitrogen), and analyzed by flow cytometry. Analysis of phospholylated-p38MAPK was performed as described (16 (link)). In brief, cells were surface stained, fixed, permeabilized, and stained with antiphospho-p38MAPK–PE (BD Biosciences, 612565) antibody. Mouse anti-IgG1–PE antibody (BD Biosciences) served as control. Cells were analyzed by flow cytometry. Phospho-p38MAPK–PE intensity was normalized to the mean fluorescence intensity of the IgG control.

The following mAbs were used in this study: rat mAbs against CD16/32 (93; eBioscience), c-Kit (2B8; eBioscience and Tonbo Biosciences), Sca-1 (E13-161.7; BioLegend), CD4 (L3T4; BD Biosciences), CD8a (53-6.72; BD Biosciences), B220 (RA3-6B2; BD Biosciences), TER-119 (TER119; Tonbo Biosciences), Gr-1 (RB6-8C5; BD Biosciences), CD34 (RAM34; eBioscience), Mac-1 (M1/70; BD Biosciences and eBioscience), Flt3 (A2F10.1; BioLegend), CD45.2 (104; BD Biosciences), CD45.1 (A20; BD Biosciences), IL7Rα (SB/119; BioLegend), CD41 (MWReg30; BD Biosciences), CD48 (HM48-1; BioLegend), CD150 (TC15-12F12.2; BioLegend), and CD45 (30-F11; BioLegend and eBioscience and BD Biosciences). A mixture of mAbs against CD4, CD8, B220, TER-119, Mac-1, and Gr-1 was used as a lineage marker (lineage). A mouse anti–Ki67-Alexa Fluor 555 antibody (B56; BD Biosciences) was used to assess the cell cycle, and mouse antiphosphorylated-p38MAPK–phenylephrine (PE) antibody (36/p38; BD Biosciences) and mouse anti-IgG1–PE antibody (BD Biosciences) were used to detect phosphorylated p38MAPK by intracellular flow cytometry. Annexin V–PE (556422; BD Bioscience) was used to analyze apoptosis. For immunocytochemistry, an anti-H2AX (pS139) antibody (N1-431; BD Bioscience), a rabbit anti-53BP1 polyclonal antibody (NB100-304; Novus Biologicals), and Alexa Fluor 555–conjugated anti-rabbit IgG (A-21428; Invitrogen) were used.