This antibody was evaluated in a study of commercially available antibodies against rat alpha7 Nicotinic Acetylcholine Receptors (α7 nAChR). Seven antibodies were included in the study and tested in the applications Western blotting and fluorescent immunocytochemistry.
Both tests were performed using the rat pituitary cell line GH4C1, which has no endogenous expression of α7 nAChR. For immunofluorescence, antibody staining of native cells was compared with staining of cells transfected with rat α7 nAChR cDNA. Expression of α7 nAChR in GH4C1 was confirmed by RT-PCR. The same two cell-lines were used for Western blotting, together with GH4C1 cells transfected with C-terminally GFP tagged α7 nAChR or GFP.
This antibody (Lot No D2214) did not recognize rat α7 nAChR in any of the two applications. In Western blotting multiple background bands were detected, but no differences were observed between transfected and untransfected cells.
In immunocytochemistry signals of similar intensities were observed in transfected and control cells.
For further information see: Evaluating Commercially Available Antibodies for Rat alpha7 Nicotinic Acetylcholine Receptors
Garg and Loring, Journal of Histochemistry and Cytochemistry 2017. Vol. 65(9) 499-512 https://www.ncbi.nlm.nih.gov/pubmed/28763248
On May 8, 2018 Brijesh Garg, Northeastern University, Boston wrote:
This antibody was evaluated in a study of commercially available antibodies against rat alpha7 Nicotinic Acetylcholine Receptors (α7 nAChR). Seven antibodies were included in the study and tested in the applications Western blotting and fluorescent immunocytochemistry.
Both tests were performed using the rat pituitary cell line GH4C1, which has no endogenous expression of α7 nAChR. For immunofluorescence, antibody staining of native cells was compared with staining of cells transfected with rat α7 nAChR cDNA. Expression of α7 nAChR in GH4C1 was confirmed by RT-PCR. The same two cell-lines were used for Western blotting, together with GH4C1 cells transfected with C-terminally GFP tagged α7 nAChR or GFP.
This antibody (Lot No D2214) did not recognize rat α7 nAChR in any of the two applications. In Western blotting multiple background bands were detected, but no differences were observed between transfected and untransfected cells.
In immunocytochemistry signals of similar intensities were observed in transfected and control cells.
For further information see: Evaluating Commercially Available Antibodies for Rat alpha7 Nicotinic Acetylcholine Receptors
Garg and Loring, Journal of Histochemistry and Cytochemistry 2017. Vol. 65(9) 499-512 https://www.ncbi.nlm.nih.gov/pubmed/28763248