American Society of Hematology

Fig. 3.

$Fig. 3. Subcellular fractionation of nitrogen-cavitated neutrophils on 3-layer Percoll gradient. / After nitrogen cavitation, 10 mL postnuclear supernatant was applied on top of a 3-layer Percoll gradient. After centrifugation, the gradients were fractionated by aspiration from the bottom of the tube in 1-mL fractions. Fractions 1 to 6, 7 to 12, 13 to 18, and 19 to 24 were pooled in 4 distinct populations named 1, 2, 3, and 4, respectively. These pooled fractions were assayed for myeloperoxidase (A,E), lactoferrin (B,F), gelatinase (C,G), and albumin (D,H) by immunologic assays (ELISA; A-D) and by immunoblotting (E-H). In both markers assay, the pooled fractions 1, 2, 3, and 4 were identified as α-, β1-, β2-, and γ-band, respectively. In E to H, the positions and sizes of the molecular weight markers are shown.$

Subcellular fractionation of nitrogen-cavitated neutrophils on 3-layer Percoll gradient.

After nitrogen cavitation, 10 mL postnuclear supernatant was applied on top of a 3-layer Percoll gradient. After centrifugation, the gradients were fractionated by aspiration from the bottom of the tube in 1-mL fractions. Fractions 1 to 6, 7 to 12, 13 to 18, and 19 to 24 were pooled in 4 distinct populations named 1, 2, 3, and 4, respectively. These pooled fractions were assayed for myeloperoxidase (A,E), lactoferrin (B,F), gelatinase (C,G), and albumin (D,H) by immunologic assays (ELISA; A-D) and by immunoblotting (E-H). In both markers assay, the pooled fractions 1, 2, 3, and 4 were identified as α-, β1-, β2-, and γ-band, respectively. In E to H, the positions and sizes of the molecular weight markers are shown.

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