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 2630253) resulted in an intense immunofluorescence signal of α7 nAChR transfected cells. The staining of untransfected control cells was of varying intensity, however the difference between control and transfected cells was clear.
In Western blotting a band of expected size was detected in α7 nAChR cells, but no signal was seen in α7 nAChR-GFP cells. In addition, multiple background bands were detected in all 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 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 2630253) resulted in an intense immunofluorescence signal of α7 nAChR transfected cells. The staining of untransfected control cells was of varying intensity, however the difference between control and transfected cells was clear.
In Western blotting a band of expected size was detected in α7 nAChR cells, but no signal was seen in α7 nAChR-GFP cells. In addition, multiple background bands were detected in all 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