American Society of Hematology

Fig. 3.

$Fig. 3. The 65- to 70-kd molecule is a free fragment of CD18 unassociated with CD11 chains. / (A) PMA-activated PBLs were lysed in 1% n-dodecyl β-d-maltoside and subjected to BNE followed by the second-dimension SDS-PAGE, electroblotting, and immunoperoxidase detection with a standard CD18 mAb MEM-48. The numbered spots correspond to (1) CD18 molecules present in the noncovalent integrin heterodimers, (2) free intracellular precursor forms of CD18 described earlier,15 and (3) free 65- to 70-kd CD18 fragment. It should be noted that under the conditions of BNE the noncovalent integrin heterodimeric complexes are preserved. Positions of molecular weight standards in both dimensions are indicated. (B) Detergent lysates of PMA-activated PBLs were passed through immunosorbents based on immobilized mAbs to the indicated antigens: CD18 (MEM-48), MEM-148, MHC class I (MEM-135; negative control), and the unbound fractions or whole cell lysate (None) were subjected to SDS-PAGE and Western blotting using MEM-148; only the relevant 50- to 80-kd region of the blot is shown.$

The 65- to 70-kd molecule is a free fragment of CD18 unassociated with CD11 chains.

(A) PMA-activated PBLs were lysed in 1% n-dodecyl β-d-maltoside and subjected to BNE followed by the second-dimension SDS-PAGE, electroblotting, and immunoperoxidase detection with a standard CD18 mAb MEM-48. The numbered spots correspond to (1) CD18 molecules present in the noncovalent integrin heterodimers, (2) free intracellular precursor forms of CD18 described earlier,15 and (3) free 65- to 70-kd CD18 fragment. It should be noted that under the conditions of BNE the noncovalent integrin heterodimeric complexes are preserved. Positions of molecular weight standards in both dimensions are indicated. (B) Detergent lysates of PMA-activated PBLs were passed through immunosorbents based on immobilized mAbs to the indicated antigens: CD18 (MEM-48), MEM-148, MHC class I (MEM-135; negative control), and the unbound fractions or whole cell lysate (None) were subjected to SDS-PAGE and Western blotting using MEM-148; only the relevant 50- to 80-kd region of the blot is shown.

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