A sodium dodecyl sulfate-gradient gel electrophoresis system that separates polypeptides in the molecular weight range of 1500 to 100,000

Ferritin was isolated from the livers and brains of two groups of rats, one of which was fed aluminum chloride (100 ,uM) for 1 year in the drinking water. Brain tissue contained about one-third ofthe amount offerritin found in the liver. While brain ferritin from normal rats contained 42.1 ± 14.3 mol of aluminum, that from the aluminum-fed group contained 115.4 ± 48.3 mol of aluminum per mol of ferritin. Liver ferritin from both groups contained similar amounts of both aluminum and iron, and the amounts were less than that found associated with brain ferritin. Ferritin isolated from the brains of patients who died of Alzheimer disease contained more aluminum and more iron than that from age-matched controls. Human brain ferritin is composed of two types of subunits-about 70% heavy chain (Mr, 22,000) and 30% light chain (Mr,19,500). The isoelectric focusing pattern of human brain ferritin was considerably different from that of human liver. Only 5 of the 20 brain ferritin bands migrated similarly to the acidic isoferritins from the liver, and the major component of brain ferritin, representing 30% of the total ferritin, had a pI of 8.0.

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“…21. The amount of protein isolated was determined by the method of Lowry with the appropriate correction factors (3).…”

Section: Methodsmentioning

confidence: 99%

Ferritin: isolation of aluminum-ferritin complex from brain.

Fleming

Joshi

1987

Proc. Natl. Acad. Sci. U.S.A.

137

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“…However, below 20 kDa this excellent multiphasic buffer system shows insufficient resolving power [6][7][8]. Accordingly, to achieve better separation of small proteins from SDS, some authors enhance the effective mobility of glycine by using base AMPD and raising the pH value to 8.96 in the separating gel [7,9]. With Tricine instead of glycine in the trailing phase, Schägger and von Jagow [8] developed an SDS-PAGE system that allows separation of proteins in the range between 5 and 30 kDa.…”

Section: Resultsmentioning

confidence: 99%

Ultrathin‐layer sodium dodecyl sulfate disc electrophoresis of proteins in the range from 10 to 220 kDa in homogeneous, low‐concentrated polyacrylamide gels

2003

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This study describes an ultrathin-layer sodium dodecyl sulfate (SDS) disc electrophoresis in polyacrylamide gels of a thickness of only 150 microm. By use of 2-amino-2-methyl-1,3-propanediol/glycine instead of traditional Tris/HCl buffer in the resolving phase of the gel, proteins with a wide range of molecular sizes (10 kDa to over 220 kDa) are separated in unusually low-concentrated gels (4%T, 3.3%C). 2-Amino-2-methyl-1,3-propanediol in the resolving part of the gel contributes to stabilization of the pH value at 8.8, while glycine improves destacking as well as separation of small proteins from the bulk of stacked SDS. This method combines both the advantages of conventional slab-gel electrophoresis and capillary gel electrophoresis. It is easy to apply and well suited for all further miniaturization attempts.

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“…Gel electrophoresis was carried out according to [8], and silver staining as described in [9]. Toxin concentrations were estimated using a molar extinction coefficent of 20 mM_' .…”

Section: Purification Of the Toxinmentioning

confidence: 99%

Purification and characterization of a β‐toxin from the venom of the African scorpion Leiurus quinquestriatus

1987

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The venom of the African scorpion Leiurus quinquestriatus was subjected to high-performance ion-exchange chromatography.Among a large number ( > 25) of small proteins and other substances, a protein component of approx. 6500 Da was purified. The effect of this toxin was tested on single myelinated nerve fibres of the frog Rana esculenta. Toxin concentrations less than IO nM produced clear effects. Activation rather than inactivation of the voltage-dependent sodium channel was strongly affected. Thus, this toxin from an African scorpion acts like the b-toxins present in the venom of North American scorpions.

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A sodium dodecyl sulfate-gradient gel electrophoresis system that separates polypeptides in the molecular weight range of 1500 to 100,000

Cited by 31 publications

References 16 publications

Ferritin: isolation of aluminum-ferritin complex from brain.

Ferritin: isolation of aluminum-ferritin complex from brain.

Ultrathin‐layer sodium dodecyl sulfate disc electrophoresis of proteins in the range from 10 to 220 kDa in homogeneous, low‐concentrated polyacrylamide gels

Purification and characterization of a β‐toxin from the venom of the African scorpion Leiurus quinquestriatus

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