1985
DOI: 10.1104/pp.77.4.857
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Separation of the Mg2+-ATPases from the Ca2+-Phosphatase Activity of Microsomal Membranes Prepared from Barley Roots

Abstract: Two methods for preparing membrane fractions from barley (Hordeum vulgare cv California Mariout 72) roots were compared in order to resolve reported differences between the characteristics of the plasma membrane ATPase of barley and that of other species. When microsomal membranes were prepared by a published procedure and applied to a continuous sucrose gradient, the membranes sedimented as a single broad band with a peak density of 1.16 grams per cubic centimeter (g/cm3). Activities 2) in 0.25 M sucrose, a… Show more

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Cited by 32 publications
(28 citation statements)
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“…The 10,000g supernatant was centrifuged at 100,000 rpm for 35 min in a Beckman 45 Ti rotor. The pellet was resuspended in 2 to 3 ml of suspension buffer consisting of 0.25 M sucrose and 2 mm DTT in 5 mm Pipes-KOH (pH 7.0), layered onto continuous (12) or discontinuous sucrose gradients, and centrifuged at 80,000g for 3 h. Two ml fractions were collected from the 15 to 45% (w/w) continuous gradients. The discontinuous gradients consisted of 14, 10, and 10 ml of 40, 30, and 22% (w/w) sucrose, respectively.…”
Section: Methodsmentioning
confidence: 99%
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“…The 10,000g supernatant was centrifuged at 100,000 rpm for 35 min in a Beckman 45 Ti rotor. The pellet was resuspended in 2 to 3 ml of suspension buffer consisting of 0.25 M sucrose and 2 mm DTT in 5 mm Pipes-KOH (pH 7.0), layered onto continuous (12) or discontinuous sucrose gradients, and centrifuged at 80,000g for 3 h. Two ml fractions were collected from the 15 to 45% (w/w) continuous gradients. The discontinuous gradients consisted of 14, 10, and 10 ml of 40, 30, and 22% (w/w) sucrose, respectively.…”
Section: Methodsmentioning
confidence: 99%
“…The procedures were modified from those of DuPont and Hurkman (12). The roots were excised, homogenized within 5 min of harvest, filtered through cheesecloth, centrifuged at 10,000g for 20 min in a Sorvall' GSA rotor, and the pellet discarded.…”
Section: Methodsmentioning
confidence: 99%
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