American Society of Hematology

Fig. 5.

$Fig. 5. Association of Lyn with LacCer in neutrophils but not DMSO-treated HL-60 cells. / (A) DIM fractions of neutrophils and DMSO-treated HL-60 cells were immunoprecipitated with mouse anti-LacCer antibody Huly-m13 (anti-LacCer IgM) or normal mouse IgM (normal IgM). After extraction with chloroform/methanol (2/1) solution and purification using Bond Elut-C18 packed columns, aliquots of the immunoprecipitants were also analyzed by immunoslot blotting using anti-LacCer mAb Huly-m13. (B) Immunoprecipitants with Huly m-13 shown in panel A were also analyzed by SDS-PAGE/immunoblotting using rabbit anti-Lyn IgG. (C) DIM of neutrophils or DMSO-treated HL-60 cells was immunoprecipitated with mouse anti-Lyn monoclonal IgG and subjected to HPTLC, followed by staining of LacCer with primulin. LacCer indicates LacCer standard.$

Association of Lyn with LacCer in neutrophils but not DMSO-treated HL-60 cells.

(A) DIM fractions of neutrophils and DMSO-treated HL-60 cells were immunoprecipitated with mouse anti-LacCer antibody Huly-m13 (anti-LacCer IgM) or normal mouse IgM (normal IgM). After extraction with chloroform/methanol (2/1) solution and purification using Bond Elut-C18 packed columns, aliquots of the immunoprecipitants were also analyzed by immunoslot blotting using anti-LacCer mAb Huly-m13. (B) Immunoprecipitants with Huly m-13 shown in panel A were also analyzed by SDS-PAGE/immunoblotting using rabbit anti-Lyn IgG. (C) DIM of neutrophils or DMSO-treated HL-60 cells was immunoprecipitated with mouse anti-Lyn monoclonal IgG and subjected to HPTLC, followed by staining of LacCer with primulin. LacCer indicates LacCer standard.

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