| First Author | van Endert P | Year | 2015 |
| Journal | F1000Res | Volume | 4 |
| Pages | 125 | PubMed ID | 27335633 |
| Mgi Jnum | J:240377 | Mgi Id | MGI:5883201 |
| Doi | 10.12688/f1000research.6535.1 | Citation | van Endert P, et al. (2015) Unexpected lack of specificity of a rabbit polyclonal TAP-L (ABCB9) antibody. F1000Res 4:125 |
| abstractText | 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. |
