A rapid method for desalting small volumes of solution

Neal, Michael W.; Florini, James R.

doi:10.1016/0003-2697(73)90325-4

Cited by 248 publications

(67 citation statements)

References 2 publications

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“…The supernatant was centrifuged at 6000 g for 5 min to yield the crude bacteroid pellet and a supernatant which was recentrifuged at 50 000 g for 30 min. The supernatant from this step was passed through Sephadex G25 [8] swollen in 10 mM Tris-HC1 (pH 8.0), to yield the soluble protein fraction derived from the plant material of the nodule. The crude bacteroid pellet was resuspended in 0.5 M sucrose, 50 mM Tris-HC1 (pH 8.0), 2 mM dithiothreitol, and washed twice by centrifuging at 6000 g for 5 rain and resuspending.…”

Section: Methodsmentioning

confidence: 99%

Induction of glutamate synthase during nodule development in lupin

Robertson¹,

Warburton²,

Farnden

1975

FEBS Letters

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Section: Methodsmentioning

confidence: 99%

Induction of glutamate synthase during nodule development in lupin

Robertson¹,

Warburton²,

Farnden

1975

FEBS Letters

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“…A 2 mL fraction of each extract was desalted on a G-25 desalting column (Pharmacia Fine Chemicals) using the quick centrifugation method of Neal and Florini (22). Each column was preequilibrated with 50 mM Tris-HCl (pH 7.5), containing 1 mm EDTA, prior to desalting of the extract.…”

Section: Protein Extractionmentioning

confidence: 99%

Seasonal Variation in the Antioxidant System of Eastern White Pine Needles

Anderson

Chevone²,

Hess³

1992

Plant Physiol.

254

104

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Antioxidant metabolites in eastem white pine (Pinus strobus L.) needles increased two-to fourfold from the summer to the winter season. Antioxidant enzymes in needle tissue increased between 2-and 122-fold during this same period. These seasonal changes were determined by monitoring ascorbate and glutathione concentrations and the activity of ascorbate peroxidase, glutathione reductase (GR), and superoxide dismutase. Levels of antioxidant metabolites and enzymes were observed always to be lowest during the summer, or active growing season, and highest during the winter, or dormant season. These data correlated well with the thermal kinetic window for purified GR obtained from summer needles. The minimum, apparent KmDm for two isoforms of GR (GRA and GRo) occurred at 5 and 10°C, respectively. The upper limit of the thermal kinetic window (200% of the minimum K.) for GRA and GRB was 20 and 250C, respectively, indicating that needle temperatures exceeding 250C may result in impairment of antioxidant metabolism. The needle content and kinetic properties of GR, the increased activities of other enzymes, and the high substrate concentrations observed during the winter are consistent with the protective function this pathway may provide against photooxidative, winter injury.

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“…Poly(A+) RNA was isolated using oligo-dT cellulose chromatography (10). P0 (Bio-Rad) spin columns (14) were used to remove an unidentified low mol wt substance that inhibits in vitro translation. RNA from meiotic tassels was isolated by the guanidinium/CsCl method (10).…”

Section: Rna Preparation and In Vitro Translationmentioning

confidence: 99%

Developmental Staging of Maize Microspores Reveals a Transition in Developing Microspore Proteins

Bedinger¹,

Edgerton²

1990

Plant Physiol.

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A method for the preparation of developmentally staged microspores and young pollen from maize (Zea mays) has been devised. The preparations are of sufficient purity and quantity for biochemical analysis, including the analysis of steady-state protein and RNA populations associated with each stage. A major transition in protein populations occurs during the developmental period that encompasses microspore mitosis, the asymmetric nuclear division producing the vegetative and generative nuclei. Several differences between early and late stage proteins can be detected by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two-dimensional gel electrophoresis of proteins reveals that over half of the steady-state proteins differ between the younger and older stages, either quantitative or qualitative. One protein that increases in relative abundance about fourfold is actin. In vitro translation of RNA isolated from staged microspores demonstrates changes in microspore gene expression during the same developmental period.acious remnants of the tapetal cells are deposited on the surface of the pollen grain, which desiccates and is shed from the anther upon dehiscence.The study of pollen development provides an opportunity to examine the gametophyte generation of higher plants. While recent studies have suggested that multiple molecular events are involved in the progression of microspores through the pollen developmental pathway (4,(23)(24)(25), the scope of these events was unknown. The work presented here provides a method for the preparation of developmentally staged microspores and young pollen. These preparations are of sufficient purity and quantity to allow the characterization of protein and RNA populations associated with each stage. A major transition in steady-state protein populations has been observed between the large vacuole and starch-filling stages of microsporogenesis. MATERIALS AND METHODS MaterialsMicrosporogenesis in higher plants has been well described at the genetic and cytological levels (1,2,9,11,20). The maize microspore developmental pathway can be briefly summarized as follows (1,3,6,21)

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A rapid method for desalting small volumes of solution

Cited by 248 publications

References 2 publications

Induction of glutamate synthase during nodule development in lupin

Induction of glutamate synthase during nodule development in lupin

Seasonal Variation in the Antioxidant System of Eastern White Pine Needles

Developmental Staging of Maize Microspores Reveals a Transition in Developing Microspore Proteins

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