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Query: UNIPROT:P30536 (
PBS
)
9,886
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Microvillous membrane fractions from human term placentae were prepared by differential centrifugation. Extration of membranes with
PBS
-EDTA or KCI removed soluble cytoplasmic components and serum proteins excepting trace amounts of albumin and transferrin. PAGE-
SDS
revealed 11 components in the Triton solubilized crude fraction after
PBS
-EDTA extraction. Membrane components solubilized with Triton were not fractionated by gel filtration on Bio-Gel A-50 m but DEAE-cellulose chromatography partially resolved these components. Three fractions were obtained by stepwise elution of absorbed materials using increasing concentrations of NaCl in the equilibrating buffer. These fractions were characterized using
SDS
-PAGE. The material unabsorbed to the DEAE contained two components of small molecular weight and one of them showed a positive PAS stain. The first eluted protein peak showed nine components, seven of which stained with PAS. The bulk of glycoproteins with molecular weights greater than 130 000 daltons were found in this fraction. The second eluted peak from DEAE was rich in components with molecular weights less than 42 000 daltons. Four components in this fraction were not identified in the other two ion-exchange fractions. Bands representing mobilities of albumin, transferrin and alkaline phosphatase were observed in DEAE-cellulose fractions; however, 12 components of unknown structure were revealed.
...
PMID:Characterization of solubilized microvillous membrane proteins and glycoproteins from human placental syncytiotrophoblast. 723 34
One of the more recently discovered collagens, type V (or A-B) collagen, in its native fibillar form mediates human platelet aggregation and the release of serotonin. In agreement with a recent report, it has no detectable effect on human platelets in the soluble or amorphous form. The possibility that the observed results might be due to contaminating interstitial collagens was eliminated by taking advantage of unusual solubility properties of type V collagen. Type V collagen dissolved in 0.1M acetic acid formed native-type fibrils when dialyzed against
PBS
and amorphous fibrils when dislyzed against 0.05M Tris/0.13M NaCl, pH 7.4, at 4 degrees C. Interstitial collagens remained in solution under both of these conditions. In addition, type V collagen treated with sufficient, purified synovial collagenase to digest all contaminating interstitial collagen retained its platelet-aggregating properties. The purity of type V collagen was confirmed by
SDS
-PAGE of CNBr digests. These data indicate that the quaternary structure of type V collagen is important in its recognition by platelet membranes.
...
PMID:Collagen-platelet interaction. Type V(A-B) collagen induced platelet aggregation. 735 Feb 45
Astrocyte- and neuron-enriched fractions were isolated by preparative centrifugatin from rat brain and human brain, obtained at autopsy. The brain-specific protein alpha-albumin (GFA) was studied in these fractions by quantitative immunoradiochemical and by immunocytological methods. It is concluded that morphologically intact astrocytes can be isolated not only from rat, but also from human brain. In the rat, alpha-albumin (GFA) is only found in the astrocyte-enriched fractions; this shows that detemination of alpha-albumin (GFA) is useful for the evaluation of cell separation methods for the brain. From the astrocyte-enriched fractions, four times more alpha-albumin (GFA) can be extracted when detergent (0.1%
SDS
) is used, than when an aqueous extract is made. About +/- 0.15 X 10(-6) particles were extracted in 1 ml of either
PBS
or
SDS
/
PBS
. This suggests that alpha-albumin (GFA) exists in at least two forms, one hydrosoluble, and one detergent soluble, possibly structurally bound, form.
...
PMID:Immunochemical determination and immunocytological localization of brain-specific protein alpha-albumin (GFA) in isolated astrocytes. 742 59
Circulating immune complexes (CIC) were first measured in lepromatous patients (LL) by the 125I-C1q binding assay and the polyethylene glycol (PEG) precipitation test. High levels were found by both methods (95 and 90% of positives, respectively). LL-CIC were investigated for the presence of neural antigens. CIC were precipitated in 3.5% PEG, filtered through protein A-Sepharose affinity chromatography, eluted with glycine-HCl, pH 2.8, and washed with
PBS
; fractions after CIC dissociation were studied by
SDS
-PAGE and Western blotting. The LL-CIC PEG precipitates and the glycine-HCl eluates were positive in 76 and 71% respectively against anti-myelin basic proteins (MBP) monoclonal antibody, showing a single band at 15-25 kDa similar to the one obtained incubating MBP with anti-MBP. No reaction was detected with CIC-
PBS
fractions; strips were incubated with other anti-neural antibodies such as anti-glial fibrillary acidic proteins, anti-S-100, and anti-neurofilaments, without any reactivity. Our results demonstrate that LL-CIC contain MBP as an antigen; its significance could be related to the pathogenesis of leprosy since the liberation of MBP after Mycobacterium leprae nerve damage may elicit anti-MBP autoantibodies to myelin breakdown, which reacts with peripheral nerve MBP inducing CIC formation. This mechanism may be important in demyelination and destruction of nerve in leprosy.
...
PMID:Identification of myelin basic proteins in circulating immune complexes associated with lepromatous leprosy. 751 Oct 83
Two IgM isotype monoclonal antibodies (McAb), 2H10 and 2H1, recognizing repetitive epitopes on Schistosoma egg-associated molecules were characterized and their specificities were identified in a two-site sandwich ELISA system. In consistent with the differences in immunological behaviour and specificity demonstrated with immunoelectrophoresis (IEP) and immunofluorescent antibody (IFA) techniques, absolutely negative reactions found in the heterologous detecting system with alternated capture and detecting McAbs of the two revealed a complete incompatibility giving evidences that epitopes on different molecules were recognized. Immunological liability of the target antigen SEA or SEA-TCA to the two McAbs were demonstrated on sodium periodate and trifluoroacetic acid treatment indicating the biochemical nature of these epitopes were glycosyolated molecules with apparently higher resistance to the oxidizing agent showing in 2H10 recognizing epitopes. By means of an ion-gradient Mono-Q FPLC system (Pharmacia), 2H10-reactive epitopes of SEA, being tested not so efficiently adsorbed by ConA-sepharose affinity column, was found successfully concentrated in the profile eluted with pH 8.0
PBS
at 0.2-0.4 NaCl ionic strength. Repeated trials on
SDS
-PAGE and Western blotting analysis with the reactive fractions further showed a heterogeneity of molecular weight range as well as the non-transferable property of the CHO-reactive groups.
...
PMID:[Analysis of the target epitopes recognized by two monoclonal antibodies directed to egg-associated fractions of Schistosoma japonicum]. 752 6
In the last years, latex has frequently been found to be involved in immediate hypersensitivity reactions. The first case mentioned with recurrent urticaria and laryngoedema was reported by Stern (1) in 1927. Since then, latex has also been implicated in generalized urticaria, rhinoconjunctivitis, asthma and anaphylaxis. Associated sensitization to several fruits is frequently seen in latex-allergic patients with the symptoms described above. This study was performed in seven patients (six females and one male) with hypersensitivity to latex and concomitant fruit sensitization. Six of them were healthcare personnel. The age of the patients ranged from 25-39 years, with a mean of 30 years. Prick tests and intracutaneous tests with latex (10% w/v in
PBS
), banana, chestnut, avocado, kiwi and melon were carried out. A specific histamine release test (HRT) was performed according to the fluorometric assay. Antigen-specific IgE was also performed. Latex CAP inhibition with banana and
SDS
-PAGE immunoblotting were carried out in one patient. Although in latex-allergic patients multiple sensitization to fruits may be observed, banana and avocado are those most frequently involved, followed by chestnut and melon. This is likely to be due to the presence of common antigens in these fruits and latex, as demonstrated in our study only for banana and avocado. We consider that further investigation is needed on the possible sensitization to latex in sanitary personnel reporting symptoms after fruit ingestion.
...
PMID:Fruit sensitization in patients with allergy to latex. 765 8
Extraction of adsorbed proteins from dialysis membranes that had been used during actual hemodialysis procedures was performed. The condition of extraction with
SDS
plus 2-mercaptoethanol at 95 degrees C is more efficient than with only
PBS
or with
SDS
solution without 2-mercaptoethanol at 37 degrees C.
...
PMID:Extraction of serum proteins adsorbed on the surface of dialysis membranes. 783 59
A new method for purifying IgG was adapted for use with chicken serum. Chicken serum was mixed with caprylic acid, precipitated with ammonium sulfate, and dialyzed against
PBS
. Analysis by
SDS
-PAGE and Western blot confirmed successful purification of chicken IgG. Large quantities of highly purified IgG were easily obtainable for use in immunological investigations or for labeling. This technique is rapid, inexpensive, and simpler to perform than traditional ion exchange or gel filtration chromatography.
...
PMID:Simple method to purify chicken immunoglobulin G. 793 78
Glucose-6-phosphate dehydrogenase (G6PD) was purified from rabbit reticulocytes by using a single immunoaffinity chromatographic step. Antibodies against rabbit erythrocyte G6PD were raised in a goat, purified near to homogeneity and immobilized on CarboLink gel (Pierce). Nonspecific binding sites on the matrix were saturated with myokinase from rabbit muscle. Reticulocyte lysate was directly loaded onto the column, allowed to enter the gel bed and incubated for 15 min at room temperature. The column was washed first with
PBS
and then with 1 M NaCl. The enzyme was eluted with 0.1M acetic acid in 1M NaCl. Two protein bands of about 66.2 kDa were co-eluted with the G6PD, which was however well separated in
SDS
-PAGE. The eluent destroyed the enzyme activity but the G6PD yield was higher than 90%.
...
PMID:Purification of glucose-6-phosphate dehydrogenase from rabbit reticulocytes by immunoaffinity chromatography. 805 Aug 74
Different antigen preparations, viz. excretory-secretory antigen (ES Ag), phosphate buffer saline soluble antigen (
PBS
-S Ag) and sodium dodecyl sulphate soluble antigen (
SDS
-S Ag) of M. tuberculosis (M.tb) H37Ra strain along with tuberculin purified protein derivative (PPD) were employed in stick indirect ELISA to detect IgG antibodies in sera of sputum positive pulmonary tuberculosis cases. Sera from healthy individuals and individuals with diseases other than tuberculosis (cross-reacting diseases) were used as controls. ES antigen and PPD showed higher antibody titres in tuberculosis cases (GMT-1378 each) compared to
PBS
-S Ag (GMT-454) and
SDS
-S Ag (GMT-974). Thereafter, an extensive study was done analysing higher number of sera in each group for the detection of tuberculous IgG antibodies using ES Ag and PPD. The ES Ag showed better sensitivity (87%) and specificity (85%) compared to the sensitivity (73%) and specificity (78%) achieved with PPD. The ES Ag also showed higher IgG antibody titre (GMT-1068) than PPD (GMT-721). From the present study it can be envisaged that ES Ag has high diagnostic potential in tuberculosis.
...
PMID:Detection of tuberculous IgG antibodies using Mycobacterium tuberculosis H37Ra, excretory secretory antigen and tuberculin-purified protein derivative. 807 Aug 35
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