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 L0312) 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. No immunofluorescence signal was detected in any of the tested cell lines.
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 April 29, 2018 Ralph H Loring, Department of Pharmaceutical Science, Northeastern University, Boston, Massachusetts 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 L0312) 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. No immunofluorescence signal was detected in any of the tested cell lines.
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