A semiautomated hemolysis microassay for membrane lytic proteins

John Ding E. Young; Lauren Gee Leong; Marie A. DiNome; Zanvil A. Cohn

doi:10.1016/0003-2697(86)90042-4

A semiautomated hemolysis microassay for membrane lytic proteins

John Ding E. Young^*
, Lauren Gee Leong
, Marie A. DiNome
, Zanvil A. Cohn

^*Corresponding author for this work

Rockefeller University

Research output: Contribution to journal › Journal Article › peer-review

44 Scopus citations

Abstract

A simple, rapid, semiautomated microassay for hemolysis using a microtiter plate spectrophotometric system is described. The assay relies on the differences in light scattering (turbidity) properties of intact and of lysed erythrocytes. Lysis of erythrocyte suspensions in 96-well plates is determined by absorbance at 690 nm. A linear correlation between the percentage of hemolysis and the turbidity decrease is observed, indicating that this assay may be used for both rapid screening and quantitation of the hemolytic activity. This assay allows screening of 300 samples in less than 6 min. Small samples derived from protein fractionation columns (HPLC, for example) can be rapidly screened. This assay has been used in the successful isolation of a cytolytic membranelytic protein from the granules of cloned cytotoxic T lymphocytes and NK cells.

Original language	English
Pages (from-to)	649-654
Number of pages	6
Journal	Analytical Biochemistry
Volume	154
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(86)90042-4
State	Published - 01 05 1986
Externally published	Yes

Keywords

NK cells
cytotoxic T lymphocytes
hemolysis assay
pore-forming proteins microtest plates
turbidity

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10.1016/0003-2697(86)90042-4

Cite this

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title = "A semiautomated hemolysis microassay for membrane lytic proteins",

abstract = "A simple, rapid, semiautomated microassay for hemolysis using a microtiter plate spectrophotometric system is described. The assay relies on the differences in light scattering (turbidity) properties of intact and of lysed erythrocytes. Lysis of erythrocyte suspensions in 96-well plates is determined by absorbance at 690 nm. A linear correlation between the percentage of hemolysis and the turbidity decrease is observed, indicating that this assay may be used for both rapid screening and quantitation of the hemolytic activity. This assay allows screening of 300 samples in less than 6 min. Small samples derived from protein fractionation columns (HPLC, for example) can be rapidly screened. This assay has been used in the successful isolation of a cytolytic membranelytic protein from the granules of cloned cytotoxic T lymphocytes and NK cells.",

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doi = "10.1016/0003-2697(86)90042-4",

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T1 - A semiautomated hemolysis microassay for membrane lytic proteins

AU - Young, John Ding E.

AU - Leong, Lauren Gee

AU - DiNome, Marie A.

AU - Cohn, Zanvil A.

PY - 1986/5/1

Y1 - 1986/5/1

N2 - A simple, rapid, semiautomated microassay for hemolysis using a microtiter plate spectrophotometric system is described. The assay relies on the differences in light scattering (turbidity) properties of intact and of lysed erythrocytes. Lysis of erythrocyte suspensions in 96-well plates is determined by absorbance at 690 nm. A linear correlation between the percentage of hemolysis and the turbidity decrease is observed, indicating that this assay may be used for both rapid screening and quantitation of the hemolytic activity. This assay allows screening of 300 samples in less than 6 min. Small samples derived from protein fractionation columns (HPLC, for example) can be rapidly screened. This assay has been used in the successful isolation of a cytolytic membranelytic protein from the granules of cloned cytotoxic T lymphocytes and NK cells.

AB - A simple, rapid, semiautomated microassay for hemolysis using a microtiter plate spectrophotometric system is described. The assay relies on the differences in light scattering (turbidity) properties of intact and of lysed erythrocytes. Lysis of erythrocyte suspensions in 96-well plates is determined by absorbance at 690 nm. A linear correlation between the percentage of hemolysis and the turbidity decrease is observed, indicating that this assay may be used for both rapid screening and quantitation of the hemolytic activity. This assay allows screening of 300 samples in less than 6 min. Small samples derived from protein fractionation columns (HPLC, for example) can be rapidly screened. This assay has been used in the successful isolation of a cytolytic membranelytic protein from the granules of cloned cytotoxic T lymphocytes and NK cells.

KW - NK cells

KW - cytotoxic T lymphocytes

KW - hemolysis assay

KW - pore-forming proteins microtest plates

KW - turbidity

UR - https://www.scopus.com/pages/publications/0022542254

U2 - 10.1016/0003-2697(86)90042-4

DO - 10.1016/0003-2697(86)90042-4

M3 - 文章

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AN - SCOPUS:0022542254

SN - 0003-2697

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SP - 649

EP - 654

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

A semiautomated hemolysis microassay for membrane lytic proteins

Abstract

Keywords

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