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Table 1 Antibodies used in this study

From: Sequential generation of olfactory bulb glutamatergic neurons by Neurog2-expressing precursor cells

Target protein Antigen Species Dilution Source Antibody characterization
GFP Protein isolated from Aequorea victoria Rabbit 1:500 Invitrogen Recognizes a single band at 27 kDa on western blot of GFP transgenic mouse brain. No immunostaining seen in sections not containing GFP
GFP Recombinant full length protein Chicken 1:1,000 Abcam ab13970 Recognizes a single band at 27 kDa on western blot of GFP transgenic mouse brain. No immunostaining seen in sections not containing GFP
Tbr2 Synthetic peptide from mouse Tbr2 Rabbit l 1:1,000 Abcam ab23345 The antibody recognizes a single band at 72 kDa on a western blot of whole embryonic mouse brain (manufacturer's website). No western blot staining after neutralization of antibody with Tbr2 peptide
Tbr1 Synthetic peptide from mouse Tbr1 Rabbit 1:1,000 Millipore AB9616 The antibody recognizes the 68 kDa Tbr1 protein on a western blot (manufacturer's data sheet). No staining seen in Tbr1 knock-out animal (manufacturer's personal communication)
Neurog2 Carboxyl terminus peptide of Neurogenin 2 of human origin Goat 1:100 Santa Cruz SC19233 No western blot staining after neutralization of antibody with Neurog2 peptide
Calretinin Recombinant human calretinin-22k Mouse 1:1,500 Swant 6B3 This antibody specifically recognizes the 29-kDa band on immunoblots of brain extracts. The antibody staining is absent in calretinin knock-out mice (manufacturer's website)
Calbindin recombinant rat calbindin D-28k Rabbit 1:5,000 Swant CB38 This antibody specifically recognizes the 28-kDa band on immunoblots of brain extracts. The antibody staining is absent in calretinin knock-out mice (manufacturer's website)
Tyrosine hydroxylase Tyrosine hydroxylase purified from PC12 cells Mouse 1:250 Millipore MAB318 Recognizes a protein of approximately 59 to 61 kDa by western blot and does not cross-react with dopamine-hydroxylase, phenylalanine hydroxylase, tryptophan hydroxylase, or phenyl ethanolamine-N-methyl transferase on western blots (manufacturer's website). This antibody stained only cells with the typical morphology and positioning of dopaminergic periglomerular cells in the olfactory bulb
Parvalbumin Parvalbumin purified from carp muscles Mouse 1:5,000 Swant PV235 Specifically stains the 45Ca-binding spot of parvalbumin (MW 12,000 and IEF 4.9) in a two-dimensional 'immunoblot'. No antibody staining seen in a parvalbumin knock-out mouse (manufacturer's website).
Pax6 Peptide from the carboxyl terminus of the mouse Pax6 protein Rabbit 1:250 Covance PRB-278P The antibody recognizes a single band at 49 kDa on a western blot of whole embryonic mouse brain (manufacturer's personal communication)
NeuN Purified cell nuclei from mouse brain Mouse 1:500 Millipore MAB377 Recognizes two to three bands in the 46- to 48-kDa range and possibly another band at approximately 66 kDa. Given that NeuN is defined by the antibody, it cannot be characterized. However for further discussion see [58]
HuC/D HuC/HuD neuronal protein Mouse 1:250 Invitrogen A-21271 Recognizes the Elav family members HuC, HuD and Hel-N1, which are all neuronal proteins. Anti-HuC/D monoclonal 16A11 does not recognize HuR, another Elav family member that is present in all proliferating cells
GAD65 Affinity-purified GAD from rat brain Mouse 1:1,000 Abcam AB26113 Recognizes selectively GAD65; extensively co-localizes with VGAT in GABAergic axon terminals [28]
  1. Swant (Marly, Fribourg, Switzerland)
  2. Abcam (Cambridge, County of Cambridgeshire, UK)
  3. Millipore (Billerica, Massachusetts, USA)
  4. Santa Cruz Biotechnology (Santa Cruz, California, USA)
  5. Invitrogen (Carlsbad, California, USA)
  6. Covance (Princeton, New Jersey, USA)