1986
DOI: 10.1182/blood.v67.4.1126.1126
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Evaluation by quantitative acid elution and radioimmunoassay of multiple classes of immunoglobulins and serum albumin associated with platelets in idiopathic thrombocytopenic purpura

Abstract: Immunoglobulins (Igs) and serum albumin were eluted from normal platelets and platelets from patients with idiopathic thrombocytopenic purpura (ITP) with a quantitative acid elution procedure followed by solid-phase radioimmunoassay (SPRIA). Acid elution was shown to release a reproducible fraction of platelet-associated Igs, and the amounts released per platelet were independent of the platelet concentration over a wide range of concentrations. This procedure is suitable for sensitive, reproducible, and speci… Show more

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Cited by 41 publications
(10 citation statements)
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“…One million neutrophils were washed twice in PBS and suspended in 100 pl PBS with 1% bovine serum albumin (BSA), then incubated with 100 pl of serum for 30 minutes at room temperature. After 4 washes with cold PBS, the IgG bound to these cells was eluted, using a modification of the technique described by Hotchkiss et al (24). Briefly, the washed PMNs were suspended in 700 pl PBS/BSA and transferred to a microfuge tube containing 375 pl of eluting buffer (75 mM acetic acid, 150 mM NaCI, pH 2.7).…”
Section: Methodsmentioning
confidence: 99%
“…One million neutrophils were washed twice in PBS and suspended in 100 pl PBS with 1% bovine serum albumin (BSA), then incubated with 100 pl of serum for 30 minutes at room temperature. After 4 washes with cold PBS, the IgG bound to these cells was eluted, using a modification of the technique described by Hotchkiss et al (24). Briefly, the washed PMNs were suspended in 700 pl PBS/BSA and transferred to a microfuge tube containing 375 pl of eluting buffer (75 mM acetic acid, 150 mM NaCI, pH 2.7).…”
Section: Methodsmentioning
confidence: 99%
“…After 4 washings over 1 h, the blots were incubated with horse radish peroxidase (POD)-conjugated GAH IgG and IgM (Dianova, Hamburg, FRG, dilution 1 : 4,000), washed again 4 times and developed with peroxidase substrate (20 mg 3-amino-9-ethylcarbazole, 2.5 ml N, N, dimethylformamid, 50 p1 30% H202 in 50 ml 10 mM tris, pH 8.0). After [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] min, the reactions were stopped with 0.02% NaN3 in washing buffer.…”
Section: Immunoblottingmentioning
confidence: 99%
“…The technique was adapted from the acid elution technique described by Hotchkiss et al [15]. Briefly, 3x108 platelets were incubated with approximately 50 pl antiserum for 30 rnin at 37'C, washed 3 times with 500 p10.…”
Section: Acid Elutionmentioning
confidence: 99%
See 1 more Smart Citation
“…However, detection of platelet-bound antibodies is difficult to carry out when platelet count is very low, as it is at the onset of ITP. Furthermore, elevated levels of platelet-associated immunoglobulins (and other plasmaproteins, such as albumin and C3) are also detectable in patients with nonimmune thrombocytopenia [12,14].…”
Section: Introductionmentioning
confidence: 99%