Fig. 1.
Fig. 1. Generation and analysis of rat anti-mouse CD150 monoclonal antibodies. / (A) mCD150/HA/MYC construct expressed in CHO cells used to immunize rats to generate anti-mCD150 monoclonal antibodies. The extracellular region of mCD150 was generated with PCR and cloned into theBglII and SalI sites of the pDISPLAY vector yielding a HA/MYC-tagged, membrane-anchored extracellular mCD150 (see “Materials and methods”). The mCD150-EGFP construct was generated by generating full-length cDNA of mCD150 with CD150 primers encoding anEcoR1 site (5′) and BamH1 site (3′) and cloning the PCR product into the corresponding restriction sites of pEGFP-N3 as described in “Materials and methods.” (B) Anti-CD150 staining of wild-type mouse C57Bl/6 thymocytes, but not CD150−/−thymocytes with 4 rat antimouse CD150 monoclonal antibodies. Thin-lined histograms represent control staining with pooled rat immunoglobulin alone. Bold-lined histograms represent CD150 staining.

Generation and analysis of rat anti-mouse CD150 monoclonal antibodies.

(A) mCD150/HA/MYC construct expressed in CHO cells used to immunize rats to generate anti-mCD150 monoclonal antibodies. The extracellular region of mCD150 was generated with PCR and cloned into theBglII and SalI sites of the pDISPLAY vector yielding a HA/MYC-tagged, membrane-anchored extracellular mCD150 (see “Materials and methods”). The mCD150-EGFP construct was generated by generating full-length cDNA of mCD150 with CD150 primers encoding anEcoR1 site (5′) and BamH1 site (3′) and cloning the PCR product into the corresponding restriction sites of pEGFP-N3 as described in “Materials and methods.” (B) Anti-CD150 staining of wild-type mouse C57Bl/6 thymocytes, but not CD150−/−thymocytes with 4 rat antimouse CD150 monoclonal antibodies. Thin-lined histograms represent control staining with pooled rat immunoglobulin alone. Bold-lined histograms represent CD150 staining.

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