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Pooled medium concentrated 12-fold by dialysis and freeze-drying was assayed for sulphated glycosaminoglycans, using shark cartilage chondroitin sulphate as standards.
The analysis of culture medium concentrated by centrifugation in CsCl 1.5 g/mL, in the presence or absence of UTP, indicated that fractions enriched in apomucin 5AC contained carbohydrates like sialic acid and 5(d)), with a similar pattern distribution and immunoreactive intensity– 5(d)).
Groups of transparent (i.e. very thin) nanoleaves are discernible in the case of Sample 4 processed in the medium concentrated solution (0.43 M LiOH).
For analysis of proMMP-2 activation, conditioned medium was collected after 24 hours in serum-free medium, concentrated, and analyzed for gelatinase activity by zymography.
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For Western blot analysis of activated u-PA in conditioned culture medium, medium was concentrated 40-fold using centricon 10 ultrafiltration tubes (Millipore, Ireland).
For detection of amphiregulin (AR) shed into cell culture medium, we concentrated the medium 10X using Amicon Centriplus filters with a cutoff of 3 kDa as previously described [4].
For analysis of TGFBI protein level, conditioned medium was collected and the protein in the medium was concentrated by adding SP sepharose at the proportion of 25 μl/ml medium (Amersham, Arlington Heights, IL) and eluted with 1× SDS loading buffer.
This experiment was designed to examine the performance of Holstein-Friesian (HF) and Jersey × Holstein-Friesian (J × HF) cows within a high concentrate input total confinement system (CON) and a medium concentrate input grazing system (GRZ).
Treatments were: Low concentrate (LC), where each cow received 4 kg concentrate and 10 kg dry matter (DM) pasture per day; medium concentrate (MC), where each cow received 7 kg concentrate and 7 kg DM pasture per day; high concentrate (HC), where each cow received 10 kg concentrate and 5 kg DM pasture per day.
A low concentrate (LC) TMR (80% CS, 20% concentrate; 12.4% CP, 35% NDF), a medium concentrate (MC) TMR (60% CS, 40% concentrate; 12.3% CP. 28% NDF), and a high concentrate (HC TMRR (40% CS, 60% concentrate; 12.6% CP, 25% NDF), were fed once per day.
This 2 × 2 factorial design experiment was conducted to compare the performance of spring-calving Holstein dairy cows (HOL, n = 34) with Swedish Red × Jersey/Holstein crossbred (SR × J/HOL, n = 34) dairy cows within low and medium concentrate input grassland-based dairy systems.
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