scholarly journals Polymerization of proteins with glutaraldehyde. Soluble molecular-weight markers

1973 ◽  
Vol 135 (4) ◽  
pp. 867-873 ◽  
Author(s):  
John W. Payne
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Slight Variation ◽  
Protein Markers ◽  
Molecular Weight Range ◽  
Soluble Polymers ◽  
Wide Range ◽  
Weight Protein ◽  
Covalently Linked

Glutaraldehyde is well known for its ability to react with proteins and to produce insoluble cross-linked aggregates. In contrast with this situation, conditions are described here which yield covalently linked soluble protein oligomers. The procedure is applicable to a wide range of proteins, and by slight variation in the reaction conditions, soluble polymers in the molecular weight range 3×104−2×107were produced. The products are valuable aṡ molecular-weight markers, e.g. in sodium dodecyl sulphate–polyacrylamide-gel electrophoresis. The inherent similarities of these oligomers make them superior to commercial molecular-weight protein markers, which may have marked differences in composition and charge.

scholarly journals Purification and characterization of subcomponent C1q of the first component of mouse complement

1981 ◽  
Vol 193 (2) ◽  
pp. 621-629 ◽  
Author(s):  
K Yonemasu ◽  
T Sasaki
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Exchange Chromatography ◽  
Polyacrylamide Gel ◽  
Molecular Weight Range ◽  
Covalently Linked

1. Mouse C1q, a subcomponent of the first component of complement, has been purified in a highly haemolytically active form by a combination of precipitation with EGTA, ion-exchange chromatography and gel filtration. Yields ranged from 3 to 5 mg/200 ml of serum, and the activity of final preparations was in the range of 2 × 10(13)-4 × 10(13) C1q effective molecules/mg. 2. The molecular weight of mouse C1q was 439 500 +/- 1586, as determined by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. 3. Mouse C1q was shown to be composed of non-covalently linked subunits, all being in the molecular-weight range 45 000-46 000, and three covalently linked chains each having a molecular weight of approx. 23 000 as determined on polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate by using non-covalently and covalently linked subunits of human C1q as markers with known molecular weights calculated theoretically previously [Porter & Reid (1978) Nature (London) 275, 699-704]. 4. Mouse C1q contained hydroxyproline, hydroxylysine, a high percentage of glycine and approx. 9% carbohydrate. The absorption coefficient and nitrogen content of C1q were also determined.

scholarly journals A Comparative Study of Distribution of Protein and Cholesterol in Various Fractions of Human Semen from Infertile and Fertile Subjects

2012 ◽  
Vol 3 (3) ◽  
pp. 78-82 ◽  
Author(s):  
MS Srinivas ◽  
Vickram Sundaram ◽  
M Ramesh Pathy ◽  
TB Sridharan
Keyword(s):  
Molecular Weight ◽  
Comparative Study ◽  
Seminal Plasma ◽  
Semen Analysis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Human Semen ◽  
Exact Function ◽  
Wide Range ◽  
Significant Difference

ABSTRACT Aim To elucidate the concentration of the protein and cholesterol in different fractions of human semen from different infertile categories and comparing them with the fertile group. Materials and methods The human semen was collected from different infertile categories including oligoasthenospermia, asthenospermia, azoospermia, normospermia, oligospermia and fertile group. Immediately after collection, the semen analysis was done as per WHO standard protocols. After that, the semen was centrifuged to get the different fractions. Four main fractions were obtained, (1) spermatozoa, (2) debris or material that precipitates at 12 K rpm for 10 minutes, (3) prostasomes which was precipitated at 20K rpm for 120 minutes, (4) seminal plasma. The protein concentration was done by Lowry's method and cholesterol was estimated by diagnostic kit. Results Sodium dodecyl sulfate—polyacrylamide gel electrophoresis (SDS PAGE) was run for all the categories of semen samples for their seminal plasma and the fertility associated protein was identified. A significant difference was found in the concentration of proteins in all subfractions when compared between control and infertile categories. Almost 86% of the protein was recovered from soluble fraction. In case of azoospermia, the protein content was very low when compared with fertile group. Seminal plasma proteins were visualized by silver staining. The molecular weight of the protein bands were ranging from 6.5 to 205 kDa. The band with molecular weight around 55 kDa was found to be missing in case of oligoasthenospermia. This particular protein is said to be fertility associated protein. The content of cholesterol for different subfraction of the human semen samples from infertile and fertile samples was compared. A wide range of cholesterol was recovered from prostasomes, that too purified. Conclusion A thrive study have to be done in all the subfractions of the semen irrespective of the category of samples to know the exact function of the each subfractions in terms of protein and cholesterol distribution. How to cite this article Sundaram V, Srinivas MS, Rao KA, Pathy MR, Sridharan TB. A Comparative Study of Distribution of Protein and Cholesterol in Various Fractions of Human Semen from Infertile and Fertile Subjects. Int J Infertility Fetal Med 2012;3(3):78-82.

scholarly journals Polymeric C-terminal cross-linked material from type-I collagen. A modified method for purification, anomalous behaviour on gel filtration, molecular weight estimation, carbohydrate content and lipid content

1980 ◽  
Vol 189 (1) ◽  
pp. 111-124 ◽  
Author(s):  
N D Light ◽  
A J Bailey
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Type I Collagen ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Type I ◽  
Molecular Weight Range ◽  
Carbohydrate Analysis ◽  
Weight Range

Polymeric cross-linked C-terminal peptide material (poly-alpha 1CB6) from mature bovine tendon type-I collagen was prepared and purified by a modification of the method previously described [Light & Bailey (1980) Biochem. J. 185, 373-381]. Poly-alpha 1CB6 was shown to exhibit concentration-dependent aggregation effects on gel filtration due to interaction with a filtration medium. The material had an amino acid content that was very similar to a mixture of alpha 1CB6 and alpha 1CB5. The material was shown to be polydisperse with a mol.wt. range of 50 000-350 000, but chromatographic fractions were relatively homogeneous over this molecular weight range with respect to amino-acid composition. The heterogeneity of the material was not due to incomplete CNBr peptide cleavage, as poly-alpha 1CB6 did not contain detectable quantities of methionine. The material showed no discrete bands on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis but gave a constant blue stain throughout the molecular weight range described above. Lipid analysis showed that the partially purified material contained elevated levels of stearate when compared to the crude CNBr-digested starting material. This may indicate the specific association of a stearic-acid-rich lipid with the peptide material. On carbohydrate analysis poly-alpha 1CB6 was shown to contain only galactose and glucose at levels of 0.72 and 0.28% respectively. The carbohydrate and amino acid analyses indicated that (alpha 1CB6)2-(alpha 1CB5)1 may be the basic cross-linked structural unit of poly-alpha 1CB6)2-(alpha 1CB5)1 units, although the carbohydrate analysis indicated that the higher molecular weight oligomers may be enriched in alpha 1CB6.

scholarly journals Characterization of Kacang Goat skin Pepsin Soluble Collagen (Psc) and Their Potency as an Antioxidant

2021 ◽  
Vol 16 (2) ◽  
pp. 75-83
Author(s):  
Rina Wahyuningsih ◽  
Rusman Rusman ◽  
Nurliyani Nurliyani ◽  
Abdul Rohman ◽  
Nanung Agus Fitriyanto ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Acetic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Differential Scanning Calorimetric ◽  
Molecular Weight Range ◽  
Goat Skin ◽  
Skin Collagen ◽  
Nacl Concentration ◽  
Antioxidant Potency

Collagen have been interesting material for many utilization such as food, pharmaceutical and cosmetic in various products and target administration, consequently collagen should be prepared as well as type of application. The objective of this research is to prepare collagen from goat skin and investigate the character and their potency as an antioxidant. Kacang goat skin aged 2 years was used for collagen production. Small slice skin was extracted by curing with 0.1% (w/v) pepsin in acetic acid 0.5 M, for 24, 48, dan 72 h at 4°C. The variables observed were molecular weight by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE), microstructure using scanning electron microscope, thermal stability by differential scanning calorimetric, and the antioxidant potency through 2,2-diphenyl-1-picrylhydrazyl (DPPH) inhibition analysis. The result showed the molecular weight range from 25 kDa to 180 kDa, microstructure showed the collagen fibril crosslink, collagen start to denature at 62,28°C, highest dissolved with 1% NaCl concentration and has highest DPPH inhibition at 60 min after hydrolysis. In conclusion, kacang goat skin collagen prepared by pepsin in acetic acid.

scholarly journals Genetic Diversity of High and Low Molecular Weight Glutenin Subunits in Algerian Aegilops geniculata

2014 ◽  
Vol 42 (2) ◽  
pp. 453-459 ◽  
Author(s):  
Asma MEDOURI ◽  
Inès BELLIL ◽  
Douadi KHELIFI
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Storage Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Glutenin Subunits ◽  
Bread Making ◽  
Aegilops Geniculata ◽  
Wide Range

Aegilops geniculata Roth is an annual grass relative to cultivated wheat and is widely distributed in North Algeria. Endosperm storage proteins of wheat and its relatives, namely glutenins and gliadins, play an important role in dough properties and bread making quality. In the present study, the different alleles encoded at the four glutenin loci (Glu-M1, Glu-U1, Glu-M3 and Glu-U3) were identified from thirty five accessions of the tetraploid wild wheat A. geniculata collected in Algeria using Sodium dodecyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE). At Glu-M1 and Glu-U1 loci, encoding high molecular weight glutenin subunits (HMW-GS) or A-subunits, 15 and 12 alleles were observed respectively, including one new subunit. B-Low molecular weight glutenin subunits zone (B-LMW-GS) displayed a far greater variation, as 28 and 25 alleles were identified at loci Glu-M3 and Glu-U3 respectively. Thirty two subunits patterns were revealed at the C subunits- zone and a total of thirty four patterns resulted from the genetic combination of the two zones (B- and C-zone). The wide range of glutenin subunits variation (high molecular weight glutenin subunits and low molecular weight glutenin subunits) in this species has the potential to enhance the genetic variability for improving the quality of wheat./span>

A Comparative Study of Crosslinked and Noncrosslinked Fibrin from the Major Classes of Vertebrates

1973 ◽  
Vol 29 (02) ◽  
pp. 313-338 ◽  
Author(s):  
Martin L. Schwartz ◽  
Salvatore V. Pizzo ◽  
J Bolling Sullivan ◽  
Robert L. Hill ◽  
Patrick A. McKee
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cartilaginous Fish ◽  
Transient Species ◽  
Molecular Weight Range ◽  
Crosslinking Process ◽  
Bony Fishes

SummaryCrosslinked and noncrosslinked fibrin formed by clotting whole plasma in the presence and absence of calcium has been examined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The fibrin from 45 species of vertebrates with representatives from all classes except the Bradyodonti (Chimaeras) and a majority of the subclasses have been compared. Noncrosslinked fibrin from the mammals contained subunits resembling the α, β, and γ chains of human noncrosslinked fibrin. The molecular weight of the α-chains varied greatly, the molecular weight of the β-chains varied slightly, while the molecular weight of the γ-chains was apparently invariant. Nonerosslinked avian fibrins showed clearly resolved γ-chains with a molecular weight slightly smaller than human γ-chains. Avian α- and β-chains were usually not resolved. Nonerosslinked fibrin from the reptiles and amphibians contained a clearly resolved subunit with a molecular weight similar to that of human γ-chain. In crosslinked fibrin from all of the mammals, birds, reptiles, and amphibians the γ-chain was absent and a new band which corresponded to a dimer of the γ-chain was found, while high molecular weight polymers were only found in a few species. Fibrin formed from the plasma of the bony fishes was often difficult because of the problem of fibrinolysis ; however, non crosslinked fibrin from some species of bony fish had three clearly resolved subunits while the crosslinked fibrin from all of the species examined had dimers as the predominant crosslinked forms. The noncrosslinked fibrin from the cartilaginous fish had subunits in the same molecular weight range as the other vertebrate fibrins; however, the crosslinked fibrin was unique to this class because all of the fibrin subunits were involved in the crosslinking process. Dimers appeared to be a transient species, and a large number of different high molecular weight crosslinked species were formed. The crosslinked fibrin from the hagfish contained dimers and no higher molecular weight crosslinked forms. Rapid fibrinolysis complicated the interpretation of the results from all of the classes of fish.

Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis of Proteins

2021 ◽  
Vol 2021 (12) ◽  
pp. pdb.prot102228
Author(s):  
Clara L. Kielkopf ◽  
William Bauer ◽  
Ina L. Urbatsch
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Polypeptide Chain ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Relative Size ◽  
Apparent Molecular Weight ◽  
Protein Markers ◽  
Sds Page ◽  
Dodecyl Sulfate

Most analytical electrophoreses of proteins are achieved by separation in polyacrylamide gels under conditions that ensure dissociation of proteins into individual polypeptide subunits and minimize aggregation. Most commonly, the anionic detergent sodium dodecyl sulfate (SDS) is used in combination with a reducing agent (β-mercaptoethanol or dithiothreitol) and with heating to dissociate proteins before loading onto the gel. SDS binding denatures the polypeptides and imparts a negative charge that masks their intrinsic charge. The amount of SDS bound is generally sequence-independent and proportional to molecular weight; at saturation, approximately one SDS molecule is bound per two amino acids, or ∼1.4 g of SDS per gram of polypeptide. Therefore, the migration of SDS–polypeptide complexes in an electric field is proportional to the relative size of the polypeptide chain, and its molecular weight can be estimated by comparison to protein markers of known molecular weight. However, hydrophobicity, highly charged sequences, and certain posttranslational modifications such as glycosylation or phosphorylation may also influence migration. Thus, the apparent molecular weight of modified proteins does not always accurately reflect the mass of the polypeptide chain. This protocol describes preparation and running of SDS-PAGE gels, followed by staining to detect proteins using Coomassie Brilliant Blue. Finally, the stained SDS-PAGE gel may be scanned to an image or preserved by drying.

Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

1992 ◽  
Vol 68 (05) ◽  
pp. 534-538 ◽  
Author(s):  
Nobuhiko Yoshida ◽  
Shingi Imaoka ◽  
Hajime Hirata ◽  
Michio Matsuda ◽  
Shinji Asakura
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Tetraacetic Acid ◽  
Fibrin Monomer ◽  
Sds Page ◽  
Thrombotic Tendency ◽  
Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

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