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Example sentences for: affinity-purified
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A band for RIIβ was detected in contrast to a report of no RIIβ detection in soleus or extensor digitorum longus muscles [ 18 ] , which might be explained by the presence of nerve fibers or adipocytes wherein RIIβ is prevalent [ 27 ] . The blot for the affinity-purified D-AKAP1 antibody is shown because the protein-A purified antibody displayed additional, unexplained bands; however, no difference in fluorescence labeling was observed when using antibodies purified either way.
The rabbit antiserum obtained was affinity-purified by passage over a Sulfolink column (Pierce, Rockford, IL, USA) to which disulfide linkage of the peptide had been used to immobilize the peptide.
Antibodies: SKP2 (Zymed 32-3300, 1:250 for immunoblotting), SKP2 (Santa Cruz sc-7164, 1:500 for immunofluorescence staining), p27 (Transduction Laboratories K25030, 1:5000 for immunoblotting, 1:1000 for immunofluorescence staining), cyclin A (Neomarkers Ab-6, MS-1061, 1:100 for immunoblotting), cyclin E (Neomarkers Ab-1, RB-012, 2 ug per immunoprecipitation) cyclin E (Neomarkers Ab-2, MS-870, 1:200 for immunoblotting), HRT1 (affinity-purified rabbit polyclonal, gift from R. Deshaies, 1:1000 for immunoblotting), CUL1 (Zymed 71-8700, 1:250 for immunoblotting), SKP1 (Transduction laboratories 610530, 1:5000 for immunoblotting), JAB1/CSN5 (GeneTex MS-Jab11-PX1, 1:1000 for immunoblotting), RB1 (Pharmingen 554136, 1:500 for immunoblotting), CDK2 (Santo Cruz Biotechnology sc-163, 2 ug per immunoprecipitation, 1:500 for immunoblotting), Tubulin (Sigma T5168, 1:2000 for immunoblotting).
Following incubation in blocking buffer (10 mM Tris-HCl, pH 7.5, containing 10% non-fat dry milk, 100 mM NaCl, and 0.1% Tween 20), the membranes were probed with an affinity-purified rabbit polyclonal IgG directed against caspase-8 (Caspase-8 p20, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), 1:200 dilution in blocking buffer, overnight at 4°C.
As shown in Figure 4, affinity-purified mactinin antisera reacted with several 30-40 kDa proteins, including 31 kDa mactinin in each of the samples tested.