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Specificity: 2A1 exhibited polyreactive recognition of peptide 100-119 from the alpha subunit and peptide 100-119 frmo the beta subunit. Moreover, 2A1 bound Hb and the Hb-haptoglobin complex with equal efficiency, implying different binding sites on Hb for haptoglobin and 2A1. 2A1 was also found to distinguish between Hb and metHb.
Preparation: Murine hybridomas were generated using spleen cells from aging (8-10 month old) NZB/W F1 animals. Hybridomas secreting antibodies reactive to murine hemoglobin were subcloned by limiting dilution and antibody isotypes determined using commercial reagents. Eight-week-old NZB/W F1 were immunized subcutaneously with 50 mg mHb antigen emulsified in Complete Fruends Adjuvant.
The human alpha globin gene cluster located on chromosome 16 spans about 30 kb and includes seven loci: 5'- zeta - pseudozeta - mu - pseudoalpha-1 - alpha-2 - alpha-1 - theta - 3'. The alpha-2 (HBA2) and alpha-1 (HBA1) coding sequences are identical. These genes differ slightly over the 5' untranslated regions and the introns, but they differ significantly over the 3' untranslated regions. Two alpha chains plus two beta chains constitute HbA, which in normal adult life comprises about 97% of the total hemoglobin; alpha chains combine with delta chains to constitute HbA-2, which with HbF (fetal hemoglobin) makes up the remaining 3% of adult hemoglobin. Alpha thalassemias result from deletions of each of the alpha genes as well as deletions of both HBA2 and HBA1; some nondeletion alpha thalassemias have also been reported.
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