American Society of Hematology

Figure 4.

$Figure 4. Major erythrocyte raft components include band 3, the flotillins, and stomatin. Detergent-resistant membrane fractions were prepared from normal human erythrocytes. Membranes were purified, extracted in 1% cold Tx-100, and subjected to sucrose density gradient centrifugation for 18 hours as described in “Materials and methods.” The raft-containing fraction was dialyzed, dried, TCA precipitated, dried, and resuspended. The purified fraction was separated by reducing SDS-PAGE and stained with collodial Coomassie or silver. Structural analyses were performed on excised Coomassie-stained protein bands using a nanocapillary reverse-phase column coupled to a ThermoFinnigan LCQ Classic quadrupole ion trap mass spectrometer. Resulting masses and MS/MS data were searched against the nonredundant NCBI database to make the listed protein predictions. *Tentative predictions.$

Major erythrocyte raft components include band 3, the flotillins, and stomatin. Detergent-resistant membrane fractions were prepared from normal human erythrocytes. Membranes were purified, extracted in 1% cold Tx-100, and subjected to sucrose density gradient centrifugation for 18 hours as described in “Materials and methods.” The raft-containing fraction was dialyzed, dried, TCA precipitated, dried, and resuspended. The purified fraction was separated by reducing SDS-PAGE and stained with collodial Coomassie or silver. Structural analyses were performed on excised Coomassie-stained protein bands using a nanocapillary reverse-phase column coupled to a ThermoFinnigan LCQ Classic quadrupole ion trap mass spectrometer. Resulting masses and MS/MS data were searched against the nonredundant NCBI database to make the listed protein predictions. *Tentative predictions.

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