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 : 1000
rabbit (M, AB1504); 1 : 10,000
Secondary Ab:
donkey (MP, A11058; AF594); 1 : 500
donkey (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 : 2500
rabbit (C, AB1506); 1 : 2500
Secondary Ab:
donkey (MP, A11055; AF488); 1 : 1000
donkey (MP, A21207; AF594); 1 : 1000
GluA (GluR4) and Fos ()
Fos (red)
GluA4 receptor subunit (green)
Primary Ab:
goat (SC, sc-52G); 1 : 1000
rabbit (M, AB1508); 1 : 5000
Secondary Ab:
donkey (MP, A11058; AF594); 1 : 500
donkey (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 : 1000
mouse (M, MAB379); 1 : 5,000
Secondary Ab:
donkey (MP, A21206; AF488); 1 : 500
donkey (MP, A21203; AF594); 1 : 500
GluK4 (KA1) and Fos ()
Fos (green)
GluK4 receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 2000
rabbit (SC, sc-25700); 1 : 100
Secondary Ab:
donkey (I, A11055; AF488); 1 : 500
donkey (I, A21207; AF594); 1 : 1000
GluK5 (KA2) and Fos ()
Fos (green)
GluK5 receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 2000
rabbit (SC, sc-25701); 1 : 200
Secondary Ab:
donkey (I, A11055; AF488); 1 : 1000
donkey (I, A21207; AF594); 1 : 1000
NMDA
GluN1 (NR1, NMDAζ1) and Fos ()
Fos (green)
GluN1 receptor subunit (red)
Primary Ab:
rabbit (SC, sc-52); 1 : 5000
goat (SC, sc-1467); 1 : 700
Secondary Ab:
donkey (I, A21206; AF488); 1 : 1000
donkey (I, A11058; AF594); 1 : 1500
GluN2A (NR2a, NMDAε1) and Fos ()
Fos (green)
GluN2A receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 2000
rabbit (SC, sc-9056); 1 : 200
Secondary Ab:
donkey (I, A11055; AF488); 1 : 1000
donkey (I, A21207; AF594); 1 : 1000
GluN2B (NR2b, NMDAε2) and Fos ()
Fos (green)
GluN2B receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 2000
rabbit (SC, sc-9057); 1 : 500
Secondary Ab:
donkey (I, A11055; AF488); 1 : 1000
donkey (I, A21207; AF594); 1 : 1000
GluN2C (NR2c, NMDAε3) and Fos ()
Fos (green)
GluN2C receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 1000
rabbit (SC, sc-50437); 1 : 200
Secondary Ab:
donkey (I, A11055; AF488); 1 : 1000
donkey (I, A21207; AF594); 1 : 750
GluN2D (NR2d, NMDAε4) and Fos ()
Fos (green)
GluN2D receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 500
rabbit (SC, sc-10727); 1 : 100
Secondary Ab:
donkey (I, A11055; AF488); 1 : 1000
donkey (I, A21207; AF594); 1 : 1000
GluN3B (NR3B) and Fos ()
Fos (green)
GluN3B receptor subunit (red)
Primary Ab:
goat (SC, sc-52G); 1 : 1000
rabbit (SC, sc-50474); 1 : 150
Secondary Ab:
donkey (I, A11055; AF488); 1 : 1000
donkey (I, A21207; AF594); 1 : 1000
Substance P
NK1 and Fos ()
Fos (green)
NK1 receptor (red)
Primary Ab:
rabbit (SC, sc-52); 1 : 1000
guinea pig (M, AB15810); 1 : 2,000
Secondary Ab:
donkey (MP, A21206; AF488); 1 : 500
goat (MP, A11076; AF594); 1 : 500
Note. Glutamate receptor subunit names are consistent with IUBCP nomenclature, with common receptor subunit names in parentheses [14].