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. 1997 May 19;137(4):925–937. doi: 10.1083/jcb.137.4.925

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Chicken gizzard homogenates were separated by 10% SDS-PAGE, transferred to nitrocellulose, and probed with antibodies to either SMαA (lanes a and b) or the 1E12 antigen (lanes c and d). Gizzard homogenates were prepared by extraction in TBS alone (lanes a and c) or reextraction of the TBS-insoluble pellet in 6 M urea/TBS (lanes b and d). Immunoreactivity was determined by enhanced chemiluminescence techniques. 1E12 ascites were incubated with 0.5% SDS before incubation with the membrane and used at a final dilution of 1:1,000. 1E12 does not immunoreact with actin, but is strongly immunoreactive with a band migrating at M _r = 100,000 (arrow). In addition, a doublet migrating ahead of actin on the gel is often observed to be immunoreactive (lane d).