Primary antibody sources, concentrations used, and validation information
| Antibody name (clone/designation) | Source/citation | Concentration | Validation |
|---|---|---|---|
| TRPM1 (545H5)Ms monoclonal | Agosto et al. (2014, 2018) | 5 μg/ml, tissue 2 μg/ml, cells | WB and IF of Trpm1 KO mouse retina (Agosto et al., 2014, 2018) |
| mGluR6 (366)Ms monoclonal | Agosto et al. (2018); this study | 5 μg/ml | IF of Grm6nob3 mouse retina (this study; Fig. 4) |
| Na+/K+ ATPase (H-300)Rb polyclonal | Santa Cruz Biotechnology #sc-28800, RRID:AB_2290063 | 4 μg/ml | Expected pattern of plasma membrane staining (Robillard et al., 2012; Han et al., 2013; Iwai-Takekoshi et al., 2016) |
| PKCαRb polyclonal | Cell Signaling #2056, RRID: AB_2284227 | 1:200 | WB of macrophages from PKCα KO mouse (Koh et al., 2017); characteristic labeling of rod BPCs in mouse retina (Lu et al., 2013; Ribic et al., 2014; this study; Fig. 1) |
| 1D4 tagMs monoclonal | Molday and MacKenzie (1983); Mackenzie et al. (1984); Hodges et al. (1988) | 1 μg/ml | WB of rhodopsin KO mouse retina (Concepcion and Chen, 2010); specificity for recombinant tagged proteins (Wong et al., 2009; Agosto et al., 2014; Huynh et al., 2016) |
| HA tag (F-7)Ms monoclonal | Santa Cruz Biotechnology, #sc-7392, RRID: AB_627809 | 0.4 μg/ml | Specificity for recombinant tagged proteins (Zhao et al., 2017; Consalvi et al., 2017) |
| Ribeye A-domainRb polyclonal | Synaptic Systems, #192-103, RRID: AB_2086775 | 1 μg/ml | Correct size band on WB of retina tissue, compared to other antibodies detecting ribeye (Hubler et al., 2012); characteristic labeling of synaptic ribbon (Regus-Leidig et al., 2010; Anastassov et al., 2017; this study; Fig. 4) |
| BiP/GRP78Rb polyclonal | Abcam, #ab21685, RRID: AB_2119834 | 1 μg/ml | Colocalization with Sec61β in CHO cells and mouse retina, correct size band on WB of retina tissue (this study; Fig. 5) |
| GM130 (35/GM130) Ms monoclonal | BD Biosciences, #610822, RRID: AB_398141 | 2.5 μg/ml | WB and IF of GM130 KO mouse testes (Han et al., 2017); IF of GM130 KO liver cells (Park et al., 2018) |
| Gαo (2A)Ms monoclonal | Millipore, #MAB3073, RRID: AB_94671; Li et al. (1995) | 5 μg/ml | Correct size band on WB of bovine and rat brain membranes, specificity for Gαo vs. other Gα subunits (Li et al., 1995); widely used marker for ON-BPCs (Dhingra et al., 2002; Huang et al., 2003; Xu et al., 2012; Neuillé et al., 2015); similar staining pattern as another antibody validated by IF of Gαo KO retina (Dhingra et al., 2000) |
| Gβ3 (C-16)Rb polyclonal | Santa Cruz Biotechnology, #sc-381, RRID: AB_2109618 | 5 μg/ml | WB of Gβ3 KO mouse retina (Dhingra et al., 2012); IF of Gβ3 knockdown myocytes (Macrez et al., 1999); specific staining of ON-BPCs and cone outer segments (this study; Fig. 8), similar to another antibody validated by IF of Gβ3 KO retina (Dhingra et al., 2012) |
| Gγ13Rb polyclonal | Novus, #NBP1-98572 | 5 μg/ml | Similar staining pattern as another antibody validated by IF of Gγ13 KO retina (Ramakrishnan et al., 2015) |
Ms, mouse; Rb, rabbit; WB, western blotting; IF, immunofluorescence.