Table 1.

Primary antibodies used for immunohistochemistry

AntigenHostSourceImmunogenSpecificity
GLT1Guinea pigEMD Millipore Catalog #AB1783 RRID:AB_90949Last 20 residues of (Mars et al., 2001; Chung et al., 2008; Leung et al., 2012; Schreiner et al., 2013; Schreiner et al. 2014), or a peptide from (MTI), the C terminus of rat GLT1Western blot analysis showed a band at 50–70 kDa corresponding to GLT1 (MTI) (Redecker and Pabst, 2000; Lee et al., 2014; Roberts et al., 2014; Schreiner et al., 2014).
Preabsorption with the immunogenpeptide abolished immunostaining (MTI) (Redecker and Pabst, 2000; Lee et al., 2014) or the corresponding band on the western blot (Lee et al., 2014; Roberts et al., 2014), while preincubation with a peptide from the C-terminus region of excitatory amino acid transporter 1 had no effect on the immunostaining (Redecker and Pabst, 2000).
No staining on GLT1 null mouse tissue (Schreiner et al., 2014).
ORX AGoatSanta Cruz Biotechnology Catalog #sc-8070 RRID:unavailableA peptide from the C terminus of human orexin A (MTI; residues 48–66 of the precursor peptide (Henny and Jones, 2006))Western blot analysis confirmed specificity for rat, mouse, and human orexin A (MTI).
Preabsorption with the immunogen peptide (sc-8070P) (Henny and Jones, 2006) or orexin A (Phoenix) (Glavas et al., 2008) abolished all immunostaining.
MCHRabbitPhoenix Pharmaceuticals Catalog #H-070-47 RRID:AB_10013632Full-length MCH peptide (19 amino acids)Competitive radioimmunoassay showed 100% crossreactivity with human, rat, and mouse MCH and 0% crossreactivity with orexin A and B (MTI) (Glavas et al., 2008).
Preabsorption with full-length MCH peptide (Phoenix) abolished all staining (Glavas et al., 2008).
ChATGoatEMD Millipore Catalog #AB144P RRID:AB_2079751Human placental ChAT (identical to human brain ChAT) (Bruce et al., 1985))Western blot analysis showed a band at 68–70 kDa corresponding to ChAT (MTI) (Corcoran et al., 2004).
Preabsorption with recombinant rat ChAT (Chemicon AG220) abolished staining (Llewellyn-Smith et al., 2007; Márquez-Ruiz et al., 2007; Morcuende et al., 2011).
Identical pattern of staining with another well-characterized ChAT antibody (Umbriaco et al., 1994) was confirmed with double immunofluorescence (Nickerson Poulin et al., 2006).
PVMouse monoclonal (clone PARV-19)Sigma-Aldrich Catalog #P3088 RRID:AB_477329Frog muscle PVWestern blot analysis showed a single band at ∼12 kDa corresponding to PV (MTI).
Preabsorption with rat muscle PV eliminated labeling in Western blots and immunostaining (Hackney et al., 2005).
Identical pattern of staining with R301 below was confirmed in our lab with double immunofluorescence.
PVRabbitDr. K.G. Baimbridge (University of British Columbia) Catalog #R301Rat muscle PVPreabsorption with rat muscle PV abolished staining (Sloviter, 1989).
Identical pattern of staining with PARV-19 above was confirmed in our lab with double immunofluorescence.
  • MTI, Manufacturer's technical information.