Rabbit igg da1e

For flow cytometric analysis, the following monoclonal antibodies (mAbs) and reagents were used: CD8a (53–6.7), CD11b (M1/70), CD44 (IM7), CD45 (30-F11), CD45.1 (A20), CD45.2 (104), B220 (RA3-6B2), Thy1.2 (30-H12), Gr-1 (RB6-8C5), rat-CD2 (OX34), Hamster IgG (HTK888), Notch1 (HMN1-12), Notch2 (HMN2-35), Notch3 (HMN3-133), Notch4 (HMN4-14), anti-Rabbit-IgG (Poly4064), mIgG1 (MOPC-21), AnnexinV and 7-AAD were purchased from BioLegend. CD25 (PC61.5), CD45 (30-F11), TER119 (TER-119), Gr-1 (RB6-8C5), CD11b (M1/70), CD11c (N418), CD19 (1D3), NK1.1 (PK136), CD3e (145–2 C11), hNGFR (ME20.4) were purchased from eBioscience. CD4 (GK1.5, RM4-5) and GATA3 (L50-823) were purchased from BD Biosciences. CD19 (1D3) was purchased from Tonbo Biosciences. TCF1 (C63D9), c-Myc (D84C12) and Rabbit-IgG (DA1E) were purchased from Cell Signaling Technology. LMO2 (EP3257) was purchased from Abcam. For intracellular staining, cells were fixed and permeabilized with Foxp3/Transcription Factor Staining Set (eBioscience) or Fixation/Permeabilization Solution Kit with BD GolgiStop (BD Biosciences) and Permeabilization Buffer Plus (BD Biosciences). Stained cells were measured with FACSCalibur (BD Biosciences) or FACSVerse (BD Biosciences). Data were analyzed using FlowJo (BD Biosciences).

Abs used in FACS analysis were as follows: CD131 (REA193) from Miltenyi Biotec; FcεRIα (MAR-1) and Ki67 (SolA15) from eBioscience; phospho-S6 ribosomal protein (Ser235/236) (D57.2.2E) and phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) from Cell Signaling Technology; CD117 (c-Kit) (ACK2), Annexin V, CD123 (5B11) and CD107a (LAMP-1) (1D4B) from BioLegend. Isotype control Abs in FACS analysis were as follows: recombinant human IgG1 (REA293) from Miltenyi Biotec; rabbit IgG (DA1E) from Cell Signaling Technology; rat IgG2a (RTK2758), rat IgG2b (RTK4530), and Armenian hamster IgG (HTK888) from BioLegend. Alexa Fluoro 647-conjugated BSA (Thermo Fisher) was used as a control reagent for Annexin V staining.