Newly formed skeletal muscle fibers are prone to false positive immunostaining by rabbit antibodies

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Reports on muscle biology and regeneration often implicate immuno(cyto/histo)chemical protein characterization using rabbit polyclonal antibodies. In this study we demonstrate that newly formed myofibers are especially prone to false positive staining by rabbit antibodies and this unwanted staining is only recognized (1) by a negative muscle tissue control that does not harbor the protein to be examined (fx. from knockout mouse) or (2) by use of a nonsense rabbit antibody that has been prepared in the same way as the antibody of interest. However, many muscle immuno(cyto/histo)chemical studies only rely on controls that reveal non-specific binding by the secondary antibody and neglect that the primary rabbit antibody itself may cause false positive staining of the muscle. We suggest that reliable immuno-based protein detection in newly formed muscle fibers at least requires a nonsense rabbit antibody and optimally a negative muscle/cell control.
OriginalsprogEngelsk
TidsskriftActa Histochemica
Vol/bind133
Udgave nummer1
Sider (fra-til)68-71
ISSN0065-1281
DOI
StatusUdgivet - 2011

Fingeraftryk

Rabbits
Muscles
Proteins

Bibliografisk note

Available online 19 September 2009

Citer dette

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title = "Newly formed skeletal muscle fibers are prone to false positive immunostaining by rabbit antibodies",
abstract = "Reports on muscle biology and regeneration often implicate immuno(cyto/histo)chemical protein characterization using rabbit polyclonal antibodies. In this study we demonstrate that newly formed myofibers are especially prone to false positive staining by rabbit antibodies and this unwanted staining is only recognized (1) by a negative muscle tissue control that does not harbor the protein to be examined (fx. from knockout mouse) or (2) by use of a nonsense rabbit antibody that has been prepared in the same way as the antibody of interest. However, many muscle immuno(cyto/histo)chemical studies only rely on controls that reveal non-specific binding by the secondary antibody and neglect that the primary rabbit antibody itself may cause false positive staining of the muscle. We suggest that reliable immuno-based protein detection in newly formed muscle fibers at least requires a nonsense rabbit antibody and optimally a negative muscle/cell control.",
author = "Andersen, {Ditte C} and Anette Kliem and Schr{\o}der, {Henrik Daa} and Jensen, {Charlotte H}",
note = "Available online 19 September 2009",
year = "2011",
doi = "10.1016/j.acthis.2009.08.001",
language = "English",
volume = "133",
pages = "68--71",
journal = "Acta Histochemica",
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}

Newly formed skeletal muscle fibers are prone to false positive immunostaining by rabbit antibodies. / Andersen, Ditte C; Kliem, Anette; Schrøder, Henrik Daa; Jensen, Charlotte H.

I: Acta Histochemica, Bind 133, Nr. 1, 2011, s. 68-71.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Newly formed skeletal muscle fibers are prone to false positive immunostaining by rabbit antibodies

AU - Andersen, Ditte C

AU - Kliem, Anette

AU - Schrøder, Henrik Daa

AU - Jensen, Charlotte H

N1 - Available online 19 September 2009

PY - 2011

Y1 - 2011

N2 - Reports on muscle biology and regeneration often implicate immuno(cyto/histo)chemical protein characterization using rabbit polyclonal antibodies. In this study we demonstrate that newly formed myofibers are especially prone to false positive staining by rabbit antibodies and this unwanted staining is only recognized (1) by a negative muscle tissue control that does not harbor the protein to be examined (fx. from knockout mouse) or (2) by use of a nonsense rabbit antibody that has been prepared in the same way as the antibody of interest. However, many muscle immuno(cyto/histo)chemical studies only rely on controls that reveal non-specific binding by the secondary antibody and neglect that the primary rabbit antibody itself may cause false positive staining of the muscle. We suggest that reliable immuno-based protein detection in newly formed muscle fibers at least requires a nonsense rabbit antibody and optimally a negative muscle/cell control.

AB - Reports on muscle biology and regeneration often implicate immuno(cyto/histo)chemical protein characterization using rabbit polyclonal antibodies. In this study we demonstrate that newly formed myofibers are especially prone to false positive staining by rabbit antibodies and this unwanted staining is only recognized (1) by a negative muscle tissue control that does not harbor the protein to be examined (fx. from knockout mouse) or (2) by use of a nonsense rabbit antibody that has been prepared in the same way as the antibody of interest. However, many muscle immuno(cyto/histo)chemical studies only rely on controls that reveal non-specific binding by the secondary antibody and neglect that the primary rabbit antibody itself may cause false positive staining of the muscle. We suggest that reliable immuno-based protein detection in newly formed muscle fibers at least requires a nonsense rabbit antibody and optimally a negative muscle/cell control.

U2 - 10.1016/j.acthis.2009.08.001

DO - 10.1016/j.acthis.2009.08.001

M3 - Journal article

C2 - 19767062

VL - 133

SP - 68

EP - 71

JO - Acta Histochemica

JF - Acta Histochemica

SN - 0065-1281

IS - 1

ER -