The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins
We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Bl...
Ausführliche Beschreibung
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E-Artikel |
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Englisch |
| Erschienen: |
1987 |
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| Umfang: |
6 Ill. 6 |
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| Reproduktion: |
Wiley InterScience Backfile Collection 1832-2000 |
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| Übergeordnetes Werk: |
in: Electrophoresis - Weinheim : Wiley-VCH, 8(1987) vom: Aug., Seite 350-355 |
| Übergeordnetes Werk: |
volume:8 ; year:1987 ; month:08 ; pages:350-355 ; extent:6 |
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| 520 | |a We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. | ||
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(DE-627)NLEJ15934249X DE-627 ger DE-627 rakwb eng The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins 1987 6 Ill. 6 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. Wiley InterScience Backfile Collection 1832-2000 Tracy, Russell P. oth Monkovic, Don oth Andrianorivo, Aurelie oth Calore, James oth Mackay, Alastair oth in Electrophoresis Weinheim : Wiley-VCH 8(1987) vom: Aug., Seite 350-355 (DE-627)NLEJ15907052X (DE-600)1475486-1 0173-0835 nnns volume:8 year:1987 month:08 pages:350-355 extent:6 http://dx.doi.org/10.1002/elps.1150080804 text/html Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-WIS GBV_NL_ARTICLE AR 8 1987 8 350-355 6 |
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(DE-627)NLEJ15934249X DE-627 ger DE-627 rakwb eng The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins 1987 6 Ill. 6 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. Wiley InterScience Backfile Collection 1832-2000 Tracy, Russell P. oth Monkovic, Don oth Andrianorivo, Aurelie oth Calore, James oth Mackay, Alastair oth in Electrophoresis Weinheim : Wiley-VCH 8(1987) vom: Aug., Seite 350-355 (DE-627)NLEJ15907052X (DE-600)1475486-1 0173-0835 nnns volume:8 year:1987 month:08 pages:350-355 extent:6 http://dx.doi.org/10.1002/elps.1150080804 text/html Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-WIS GBV_NL_ARTICLE AR 8 1987 8 350-355 6 |
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(DE-627)NLEJ15934249X DE-627 ger DE-627 rakwb eng The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins 1987 6 Ill. 6 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. Wiley InterScience Backfile Collection 1832-2000 Tracy, Russell P. oth Monkovic, Don oth Andrianorivo, Aurelie oth Calore, James oth Mackay, Alastair oth in Electrophoresis Weinheim : Wiley-VCH 8(1987) vom: Aug., Seite 350-355 (DE-627)NLEJ15907052X (DE-600)1475486-1 0173-0835 nnns volume:8 year:1987 month:08 pages:350-355 extent:6 http://dx.doi.org/10.1002/elps.1150080804 text/html Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-WIS GBV_NL_ARTICLE AR 8 1987 8 350-355 6 |
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the effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins |
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The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins |
| abstract |
We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. |
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We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. |
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We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer. |
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<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ15934249X</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20210707004256.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">070201s1987 xx |||||o 00| ||eng c</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ15934249X</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">The effect of prestaining prior to immunoreaction during electrophoretic blotting of proteins</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">1987</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="b">6 Ill.</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">6</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">We have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson, Electrophoresis, 1984, 5, 35-42) to blotting of unstained gels with Amido Black or Fast Green post-transfer staining of the nitrocellulose. Contrary to a recent report (Harper et al., Anal. Biochem., 1986, 157, 270-274), in which post-staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post-staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre-view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein-dye complex is dissociated during transfer.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="f">Wiley InterScience Backfile Collection 1832-2000</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Tracy, Russell P.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Monkovic, Don</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Andrianorivo, Aurelie</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Calore, James</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Mackay, Alastair</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">in</subfield><subfield code="t">Electrophoresis</subfield><subfield code="d">Weinheim : Wiley-VCH</subfield><subfield code="g">8(1987) vom: Aug., Seite 350-355</subfield><subfield code="w">(DE-627)NLEJ15907052X</subfield><subfield code="w">(DE-600)1475486-1</subfield><subfield code="x">0173-0835</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:8</subfield><subfield code="g">year:1987</subfield><subfield code="g">month:08</subfield><subfield code="g">pages:350-355</subfield><subfield code="g">extent:6</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">http://dx.doi.org/10.1002/elps.1150080804</subfield><subfield code="q">text/html</subfield><subfield code="z">Deutschlandweit zugänglich</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-WIS</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">8</subfield><subfield code="j">1987</subfield><subfield code="c">8</subfield><subfield code="h">350-355</subfield><subfield code="g">6</subfield></datafield></record></collection>
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