Table 2 of
Spellicy, Mol Vis 2007;
13:1866-1872.Table 2 of
Spellicy, Mol Vis 2007;
13:1866-1872.
Table 2. Antibody epitopes and reactivity
Conservation of epitopes used to create antibodies used in Western blot analyses and experimentally observed reactivity of each antibody to each isoform. The red residues in the epitope conservation column indicate non-conservation of that amino acid (n.a. indicates not available). Antibodies anti-C-IMPDH1 and anti-N-IMPDH1 were raised to unique epitopes in the C and N termini of the IMPDH1 protein, and therefore are useful in distinguishing IMPDH1 from IMPDH2. Anti-C-IMPDH1 is less specific and more sensitive, and anti-N-IMPDH1 is less sensitive but more specific [18]. Antibody anti-C-IMPDH1+2 was raised to an epitope in IMPDH2, however, experimentally we found that it recognizes both forms of IMPDH (IMPDH1 and 2) simultaneously. Antibodies which distinguish between IMPDH1 retinal isoforms are anti-exon-A-IMPDH1 and anti-exon-13b-IMPDH1. The epitopes used for anti-C-IMPDH1 and anti-exon-A-IMPDH1 were 100% conserved, hence the corresponding antibodies are likely to recognize IMPDH1 proteins from other mammals. The non-conserved amino acid in the corresponding cow Anti-N-IMPDH1 epitope likely affects the ability of this antibody to recognize IMPDH1. The variable residue in the corresponding human Anti-exon-13b-IMPDH1 epitope does not seem to affect the reactivity of the antibody in the other mammalian species tested.
Reactivity Reactivity
to to Reactivity Reactivity
canonical canonical to IMPDH1 to IMPDH1
Antibody Peptide sequence Epitope conservation IMPDH1 IMPDH2 +A+13b +13b
--------------------- -------------------- -------------------------- ---------- ---------- ---------- ----------
Anti-N-IMPDH1 GFIDFIADEVDLT Human-GFIDFIADEVDLT +++ - +++ +++
Cow-GFIDFTADEVDLT
Dog-GFIDFIADEVDLT
Mouse-GFIDFIADEVDLT
Rat-GFIDFIADEVDLT
Sheep-GFIDFIADEVDLT
Pig-n.a.
Anti-C-IMPDH1 SQKRYFSEGDKVK Human-SQKRYFSEGDKVK ++++ + +++ +++
Cow-SQKRYFSEGDKVK
Dog-SQKRYFSEGDKVK
Mouse-SQKRYFSEGDKVK
Rat-n.a.
Sheep-SQKRYFSEGDKVK
Pig-n.a.
Anti-exon-A-IMPDH1 EPGARQHPGHETAAQRYSAR Human-EPGARQHPGHETAAQRYSAR - - ++++ -
Cow-EPGARQHPGHETAAQRYSAR
Dog-EPGARQHPGHETAAQRYSAR
Mouse-EPGARQHPGHETAAQRYSAR
Rat-EPGARQHPGHETAAQRYSAR
Sheep-EPGARQHPGHETAAQRYSAR
Pig-n.a.
Anti-exon-13b-IMPDH1 YTFLPFTRSGCTED Human-YTFLPFTKSGCTED - - + +
Cow-YTFLPFTRSGCTED
Dog-YTFLPFTRSGCTED
Mouse-YTFLPFTRSGCTED
Rat-n.a. Sheep-n.a.
Pig-n.a.
Anti-C-IMPDH1+2 SQNRYFSEADKIK Human-SQNRYFSEADKIK +++ +++ +++ +++
Cow-SQNRYFSEADKIK
Dog-SQNRYFSEADKIK
Mouse-SQNRYFSEADKIK
Rat-SQNRYFSEADKIK
Sheep-n.a. Pig-n.a.
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