A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes

Mitchell G. Scott; Daniel L. Crimmins; David W. McCourt; Jeffrey J. Tarrand; Margaret C. Eyerman; Moon H. Nahm

doi:10.1016/S0006-291X(88)81290-7

A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes

Mitchell G. Scott, Daniel L. Crimmins, David W. McCourt, Jeffrey J. Tarrand, Margaret C. Eyerman, Moon H. Nahm

Division of Laboratory and Genomic Medicine

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

64 Scopus citations

Abstract

We report a simple method to obtain internal amino acid sequences from larger proteins electroblotted to polyvinyldifluoride membranes. To demonstrate this method, immunoglobulin heavy and light chains are separated by gel electrophoresis and electroblotted to the membrane. The separated chains, immobilized to the membrane, are cleaved in situ by cyanogen bromide and the resulting fragments are subsequently eluted from the membrane. The fragments are separated by gel electrophoresis, electroblotted and subjected to gas phase microsequence analysis.

Original language	English
Pages (from-to)	1353-1359
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	155
Issue number	3
DOIs	https://doi.org/10.1016/S0006-291X(88)81290-7
State	Published - Sep 30 1988

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10.1016/S0006-291X(88)81290-7

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title = "A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes",

abstract = "We report a simple method to obtain internal amino acid sequences from larger proteins electroblotted to polyvinyldifluoride membranes. To demonstrate this method, immunoglobulin heavy and light chains are separated by gel electrophoresis and electroblotted to the membrane. The separated chains, immobilized to the membrane, are cleaved in situ by cyanogen bromide and the resulting fragments are subsequently eluted from the membrane. The fragments are separated by gel electrophoresis, electroblotted and subjected to gas phase microsequence analysis.",

author = "Scott, {Mitchell G.} and Crimmins, {Daniel L.} and McCourt, {David W.} and Tarrand, {Jeffrey J.} and Eyerman, {Margaret C.} and Nahm, {Moon H.}",

note = "Funding Information: ACKNOWLEDGMENTS: Supported by grant number 1902 from The Council for Tobacco Research and by a Public Health Service grant, HL 7038. DLC would like to thank Dr. Clive Slaughter and Kim Orth (HHMI, University of Texas",

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A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes. / Scott, Mitchell G.; Crimmins, Daniel L.; McCourt, David W. et al.
In: Biochemical and Biophysical Research Communications, Vol. 155, No. 3, 30.09.1988, p. 1353-1359.

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

TY - JOUR

T1 - A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes

AU - Scott, Mitchell G.

AU - Crimmins, Daniel L.

AU - McCourt, David W.

AU - Tarrand, Jeffrey J.

AU - Eyerman, Margaret C.

AU - Nahm, Moon H.

N1 - Funding Information: ACKNOWLEDGMENTS: Supported by grant number 1902 from The Council for Tobacco Research and by a Public Health Service grant, HL 7038. DLC would like to thank Dr. Clive Slaughter and Kim Orth (HHMI, University of Texas

PY - 1988/9/30

Y1 - 1988/9/30

N2 - We report a simple method to obtain internal amino acid sequences from larger proteins electroblotted to polyvinyldifluoride membranes. To demonstrate this method, immunoglobulin heavy and light chains are separated by gel electrophoresis and electroblotted to the membrane. The separated chains, immobilized to the membrane, are cleaved in situ by cyanogen bromide and the resulting fragments are subsequently eluted from the membrane. The fragments are separated by gel electrophoresis, electroblotted and subjected to gas phase microsequence analysis.

AB - We report a simple method to obtain internal amino acid sequences from larger proteins electroblotted to polyvinyldifluoride membranes. To demonstrate this method, immunoglobulin heavy and light chains are separated by gel electrophoresis and electroblotted to the membrane. The separated chains, immobilized to the membrane, are cleaved in situ by cyanogen bromide and the resulting fragments are subsequently eluted from the membrane. The fragments are separated by gel electrophoresis, electroblotted and subjected to gas phase microsequence analysis.

UR - http://www.scopus.com/inward/record.url?scp=0023783855&partnerID=8YFLogxK

U2 - 10.1016/S0006-291X(88)81290-7

DO - 10.1016/S0006-291X(88)81290-7

M3 - Article

C2 - 3178813

AN - SCOPUS:0023783855

SN - 0006-291X

VL - 155

SP - 1353

EP - 1359

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 3

ER -

A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes

Abstract

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