Artikel ; Online: Unexpected lack of specificity of a rabbit polyclonal TAP-L (ABCB9) antibody [v1; ref status
F1000Research, Vol
indexed, http://f1000r.es/5ex]
2015 Band 4
Abstract: In this article, we describe the surprising non-specific reactivity in immunoblots of a rabbit polyclonal antibody (ref. Abcam 86222) expected to recognize the transporter associated with antigen processing like (TAP-L, ABCB9) protein. Although this ... ...
Abstract | In this article, we describe the surprising non-specific reactivity in immunoblots of a rabbit polyclonal antibody (ref. Abcam 86222) expected to recognize the transporter associated with antigen processing like (TAP-L, ABCB9) protein. Although this antibody, according to company documentation, recognizes a band with the expected molecular weight of 84 kDa in HeLa, 293T and mouse NIH3T3 whole-cell lysates, we found that this band is also present in immunoblots of TAP-L deficient bone marrow-derived dendritic cell (BMDC) whole-cell lysates in three independent replicates. We performed extensive verification by multiple PCR tests to confirm the complete absence of the ABCB9 gene in our TAP-L deficient mice. We conclude that the antibody tested cross-reacts with an unidentified protein present in TAP-L knockout cells, which coincidentally runs at the same molecular weight as TAP-L. These findings underline the pitfalls of antibody specificity testing in the absence of cells lacking expression of the target protein. |
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Schlagwörter | Antigen Processing & Recognition ; Cell Signaling ; Stem Cells & Regeneration ; Medicine ; R ; Science ; Q |
Thema/Rubrik (Code) | 570 |
Sprache | Englisch |
Erscheinungsdatum | 2015-05-01T00:00:00Z |
Verlag | F1000 Research Ltd |
Dokumenttyp | Artikel ; Online |
Datenquelle | BASE - Bielefeld Academic Search Engine (Lebenswissenschaftliche Auswahl) |
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