Reverse transcription-PCR (RT-PCR) was used to amplify either the 550 nt coding for the COOH terminus
of the N gene or the full-length open reading frame for the H gene (14).
Insulin glargine (insulin A21 Gly [right arrow] Asn, and addition of 2 arginine residues to the COOH terminus
of the B chain) had ~22% cross-reactivity with E-test TOSOH II (IRI), whereas it showed concentration-dependent cross-reactivity with the ARCHITECT insulin assay (105% at 10 mIU/L and 83% at 100 mIU/L).
RNA was extracted from infected cells by using the guanidinium acid-phenol technique (11), and reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify either the 550 nucleotides (nt) coding for the COOH terminus
of N or the full-length open reading frame for H (12).
Because different antibodies used in the respective assays show different epitope specificities, they enable detection of either specifically one species, namely A[beta]1-42 [the assay of Innogenetics (11)], or the "family' of A[beta] peptides ending with a COOH terminus
at residue 42 and beginning at different [NH.sub.2] termini (ABx-42; the assays of The Genetics Co.).
If the sequence PQQ is extended one position in the direction to the NH, terminus and three positions in the direction to the COOH terminus
, then the most often occurring heptapeptide is QPQQPFP (13 times).
1), inferring that there were important antigen determinants at both termini of the N protein but that the antigenicity at the COOH terminus
was higher than that at the N[H.sub.2] terminus.
The solid-phase antibody is directed at the N[H.sub.2] terminus of the BNP peptide, whereas the conjugate antibody is directed at the COOH terminus
. Six calibrators are used for the AxSYM BNP assay.
To confirm the presence of isoforms of CRT, we performed two-dimensional Western blotting with two different antibodies: monoclonal antibody FMC75, which was produced against recombinant human CRT; and a polyclonal antibody that was produced using synthesized peptides of the human CRT COOH terminus
(amino acids 388-400) as an immunogen.
A third recombinant insulin that is longer acting is insulin glargine (Lantus[R]; Aventis Pharmaceuticals), which differs from regular human insulin by the substitution of glycine for asparagine at position A21 and by the addition of two arginine residues to the COOH terminus
of the B chain.
This problem appeared to be solved with the advent of "intact PTH" two-site IRMAs, which typically use a solid-phase capture antibody directed against the PTH COOH terminus
[usually PTH(39-84)] and a labeled detection antibody raised against PTH(1-34) (11) (see Fig.
The peptides representing the COOH terminus
of the N protein, in particular N371 and N385, had high absorbance/cutoff value ratios with the highest positive detection rate and the lowest hydrophobicity score among all of the synthesized peptides (Fig.
Because the mutations linked to Liddle syndrome were observed in the COOH termini of the [beta] and [gamma] subunits of ENaC (18), we determined the presence of mutations in exon 13 (COOH terminus
) of the Scnn1a gene to gain some insight into the role of [alpha]-ENaC in the pathogenesis of more common forms of hypertension.