Gel Filtration of Blood Platelets: A Methodological Report

Tangen, O.; Berman, Herbert J.

doi:10.1007/978-1-4684-3231-2_13

Cited by 48 publications

(19 citation statements)

References 4 publications

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“…1-and 2-day-old platelet concentrates from the NIH Clinical Center Blood Bank (prepared in acid citrate dextrose) were centrifuged at 1,200 g for 3 min to decrease red blood cell contamination. The platelets were then gelfiltered on a 2.5 X 30-cm siliconized glass column packed to a height of 20 cm with Sepharose 2B, as described by Tangen and Berman (23). Treatment of the gel, packing the column, gel filtration, eluant flow (1 ml/min), and buffer (without albumin) were as described (23).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Studies of the human factor VIII/von Willebrand factor protein. III. Qualitative defects in von Willebrand's disease.

Gralnick¹,

Coller²,

Sultan³

1975

J. Clin. Invest.

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Section: Methodsmentioning

confidence: 99%

“…The platelets were then gelfiltered on a 2.5 X 30-cm siliconized glass column packed to a height of 20 cm with Sepharose 2B, as described by Tangen and Berman (23). Treatment of the gel, packing the column, gel filtration, eluant flow (1 ml/min), and buffer (without albumin) were as described (23). The 2 Zimmerman, T. Personal communication.…”

Section: Methodsmentioning

confidence: 99%

Studies of the human factor VIII/von Willebrand factor protein. III. Qualitative defects in von Willebrand's disease.

Gralnick¹,

Coller²,

Sultan³

1975

J. Clin. Invest.

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“…Gel-filtered platelets were prepared using a modification ofthe technique described by Tangen and Berman ( 15 ). Blood was taken on 1 /6 vol of acid citrate dextrose anticoagulant and PRP was passed through a Sepharose 2B column (Pharmacia LKB Biotechnology Inc., Uppsala, Sweden) equilibrated with Hepes buffer ( 16), pH 7.35, with the following composition (mM): 3.5 Hepes, 137 NaCl, 2.7 KCI, 1.0 MgCl2, 3.0 NaH2PO4, 5.5 glucose, 0.1% BSA (Organon Teknika, Bextel, The Netherlands).…”

Section: Preparation Ofblood Samplesmentioning

confidence: 99%

A monoclonal antibody recognizes a von Willebrand factor domain within the amino-terminal portion of the subunit that modulates the function of the glycoprotein IB- and IIB/IIIA-binding domains.

Tornai

Arnout²,

Deckmyn³

et al. 1993

J. Clin. Invest.

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We developed a monoclonal antibody, 1C1E7, against vWf that increases ristocetin-induced platelet aggregation in a dose-dependent manner and lowers the threshold concentration of ristocetin needed to obtain a full aggregatory response. The platelet aggregatory effect of asialo vWf (ASvWf) also is enhanced by 1CIE7, in the presence or absence of glycoprotein (GP) Ilb/IlIa receptor antagonism. In the presence of ristocetin, both intact 1ClE7 and its Fab fragments enhance specific binding of '25I-vWf to platelets. With 1ClE7, the intermediate and higher molecular weight multimers of vWf are preferentially bound to both GP lb and GP IIb/IIIa. Thrombin-induced 125I-vWf binding to GP IIb/Illa also is increased by 1ClE7. Maximal binding of 1C1E7 to vWf corresponds to 0.97 mol/mol vWf monomer with a Kd of 4.7 x 10-10 M. 1ClE7 reacts with a 34 /36-kD tryptic fragment (III-T4) and a 34-kD plasmic fragment (P34), which localizes the epitope between amino acid residues 1 and 272; this was confirmed by NH2-terminal amino acid sequencing. Finally, platelet aggregation by ASvWf was associated with a sharp rise in intracellular Ca2+ only in the presence of 1C1E7. An antibody-mediated conformational change of vWf may result in an improved presentation of the GP Ib-and GP IIb/ IIIa-binding domains of mainly the larger multimers; the increased density of vWf on the platelet surface leads to platelet activation. The antibody may thus recognize a domain of relevance for vWf physiology. (J. Clin. Invest. 1993. 91:273-282.)

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“…The platelet-rich plasma (PRP) was recovered and centrifuged again at 24 x g (ray 8.5 cm) for 20 min to remove residual erythrocytes and leukocytes. GFP were obtained according to Tangen and Berman [16] using their noncolloidal 214 buffer, at pH 7.4. Only those preparations in which at least 90% appeared as single platelets, under the microscope, were used for studying the spectral and immunological effects upon activation.…”

Section: Preparation Of Gel-filtered Platelets (Gfp)mentioning

confidence: 99%

Conformational responses of an arachidonate‐ and U46619‐binding haemoprotein in relation to platelet activation

Krishnan

Jamaluddin

1987

FEBS Letters

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Antibodies were raised, in rabbits, against an arachidonate-and U46619-binding protein purified from calf platelets. Spectral measurements and immunodiffusion experiments were employed to follow conformational responses of the protein in relation to platelet activation. Upon treatment with the platelet agonists, arachidonate and PGH,, as well as the common haem ligands, imidazole and CN-, the purified protein had its Soret band red-shifted, with h~hromicity, but the protein saturated with the agonists, not with the haem ligands, showed altered antigenic properties in immunodiffusion experiments. In an analogous manner activation of gel-filtered calf platelets with high concentrations of ADP and A23 187, as well as by cold, had Soret bands of extracts of sonicated platelets red-shifted, with hypochromicity; concomitantly, antigenically different conformations of the protein appeared in Triton X-100 extracts of the activated platelets. A protein immunologically related to the platelet protein was detected in Triton X-100 extracts of calf neutrophils. It is suggested that confo~ational changes of the protein induced by arachidonate or prostaglandin endoperoxides or H,O, formed in different compartments during platelet activation by different stimuli may be a biochemical mechanism of stimulus-response coupling and that similar mechanisms might operate in other cell types.

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Gel Filtration of Blood Platelets: A Methodological Report

Cited by 48 publications

References 4 publications

Studies of the human factor VIII/von Willebrand factor protein. III. Qualitative defects in von Willebrand's disease.

Studies of the human factor VIII/von Willebrand factor protein. III. Qualitative defects in von Willebrand's disease.

A monoclonal antibody recognizes a von Willebrand factor domain within the amino-terminal portion of the subunit that modulates the function of the glycoprotein IB- and IIB/IIIA-binding domains.

Conformational responses of an arachidonate‐ and U46619‐binding haemoprotein in relation to platelet activation

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