International Scholarly Research Notices

Research Article

Trigeminal Medullary Dorsal Horn Neurons Activated by Nasal Stimulation Coexpress AMPA, NMDA, and NK1 Receptors

Table 1

Primary and secondary antibody combinations. Based on availability of primary and secondary antibodies and results obtained in preliminary experiments, antibodies, fluorescent color combinations, and dilutions were changed as necessary. For catalog product numbers, C: Chemicon; I: Invitrogen; M: Millipore; MP: Molecular Probes; and SC: Santa Cruz. For fluorescent tagged secondary antibodies, AF: AlexaFluor.
AMPA  
 GluA1 (GluR1) and Fos ( )  
 Fos (red)GluA1 receptor subunit (green)
  Primary Ab:goat (SC, sc-52G); 1 : 1000rabbit (M, AB1504); 1 : 10,000
  Secondary Ab:donkey (MP, A11058; AF594); 1 : 500donkey (MP, A21206; AF488); 1 : 500
 GluA2/3 (GluR2/3) and Fos ( )  
 Fos (green)GluA2/3 receptor subunits (red)
  Primary Ab:goat (SC, sc-52G); 1 : 2500rabbit (C, AB1506); 1 : 2500
  Secondary Ab:donkey (MP, A11055; AF488); 1 : 1000donkey (MP, A21207; AF594); 1 : 1000
 GluA (GluR4) and Fos ( )  
 Fos (red)GluA4 receptor subunit (green)
  Primary Ab:goat (SC, sc-52G); 1 : 1000rabbit (M, AB1508); 1 : 5000
  Secondary Ab:donkey (MP, A11058; AF594); 1 : 500donkey (MP, A21206; AF488); 1 : 500
Kainate  
 GluK1/2/3 (GluR5/6/7) and Fos ( )  
 Fos (green)GluK1/2/3 receptor subunits (red)
  Primary Ab:rabbit (SC, sc-52); 1 : 1000mouse (M, MAB379); 1 : 5,000
  Secondary Ab:donkey (MP, A21206; AF488); 1 : 500donkey (MP, A21203; AF594); 1 : 500
 GluK4 (KA1) and Fos ( )  
 Fos (green)GluK4 receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 2000rabbit (SC, sc-25700); 1 : 100
  Secondary Ab:donkey (I, A11055; AF488); 1 : 500donkey (I, A21207; AF594); 1 : 1000
 GluK5 (KA2) and Fos ( )  
 Fos (green)GluK5 receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 2000rabbit (SC, sc-25701); 1 : 200
  Secondary Ab:donkey (I, A11055; AF488); 1 : 1000donkey (I, A21207; AF594); 1 : 1000
NMDA  
 GluN1 (NR1, NMDAζ1) and Fos ( )  
 Fos (green)GluN1 receptor subunit (red)
  Primary Ab:rabbit (SC, sc-52); 1 : 5000goat (SC, sc-1467); 1 : 700
  Secondary Ab:donkey (I, A21206; AF488); 1 : 1000donkey (I, A11058; AF594); 1 : 1500
 GluN2A (NR2a, NMDAε1) and Fos ( )  
 Fos (green)GluN2A receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 2000rabbit (SC, sc-9056); 1 : 200
  Secondary Ab:donkey (I, A11055; AF488); 1 : 1000donkey (I, A21207; AF594); 1 : 1000
 GluN2B (NR2b, NMDAε2) and Fos ( )  
 Fos (green)GluN2B receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 2000rabbit (SC, sc-9057); 1 : 500
  Secondary Ab:donkey (I, A11055; AF488); 1 : 1000donkey (I, A21207; AF594); 1 : 1000
 GluN2C (NR2c, NMDAε3) and Fos ( )  
 Fos (green)GluN2C receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 1000rabbit (SC, sc-50437); 1 : 200
  Secondary Ab:donkey (I, A11055; AF488); 1 : 1000donkey (I, A21207; AF594); 1 : 750
 GluN2D (NR2d, NMDAε4) and Fos ( )  
 Fos (green)GluN2D receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 500rabbit (SC, sc-10727); 1 : 100
  Secondary Ab:donkey (I, A11055; AF488); 1 : 1000donkey (I, A21207; AF594); 1 : 1000
 GluN3B (NR3B) and Fos ( )  
 Fos (green)GluN3B receptor subunit (red)
  Primary Ab:goat (SC, sc-52G); 1 : 1000rabbit (SC, sc-50474); 1 : 150
  Secondary Ab:donkey (I, A11055; AF488); 1 : 1000donkey (I, A21207; AF594); 1 : 1000
Substance P  
 NK1 and Fos ( )  
 Fos (green)NK1 receptor (red)
  Primary Ab:rabbit (SC, sc-52); 1 : 1000guinea pig (M, AB15810); 1 : 2,000
  Secondary Ab:donkey (MP, A21206; AF488); 1 : 500goat (MP, A11076; AF594); 1 : 500
Note. Glutamate receptor subunit names are consistent with IUBCP nomenclature, with common receptor subunit names in parentheses [14].