Animal | Age | Region | Type | Detection system | Milligrams of EAAC1 per gram total proteina |
---|---|---|---|---|---|
1–3 | 220 g | Hippoc | Whole | 125I-Prot A | 0.17 ± 0.02 (3 animals) |
1–3 | 220 g | Cerebel | Whole | 125I-Prot A | 0.03 ± 0.01 (3 animals) |
4–6 | 220 g | Hippoc | Membr | 125I-Prot A | 0.25 ± 0.01 (3 animals) |
7–9 | 8 weeks, F | Hippoc | Membr | 125I-Prot A | 0.17 ± 0.01 (3 animals) |
10–12 | 14 weeks | Hippoc | Whole | Chemilum | 0.09 ± 0.01 (3 animals) |
13–19 | 9 weeks | Hippoc | Whole | Chemilum | 0.15 ± 0.01 (7 animals) |
The amounts of EAAC1 protein in tissue from the hippocampus (Hippoc) and cerebellum (Cerebel) were measured by immunoblotting using purified preparations of EAAC1 protein (Table 1) as standards. The measurements of the levels in animals 1–9 are based on anti-C491 (Ab#237) and the first EAAC1 preparation (Table 1, sample no. 1374), while the other measurements (animal 10–19) are based on anti-C491 (Ab#555) and the two other EAAC1 preparations. Bound antibodies were detected either with iodinated protein A or with chemiluminiscence (Chemilum) as indicated. Note that the cerebellum contains less EAAC1 than the hippocampus in agreement with Figure 3. Also note that some tissue samples were homogenized in water to remove water-soluble proteins before solubilization in SDS. These are indicated (Membr) in contrast to the rest, which were solubilized directly (Whole). Water-soluble proteins represent ∼50% of the total brain proteins but do not contain any detectable amounts of EAAC1 (data not shown). The water-insoluble fraction (Membr) is therefore twofold enriched with respect to EAAC1. The age is given in weight (grams) or in weeks. All animals were males except those indicated (F).
↵aValues represent mean ± SEM.