Specific cross-reactivity of antibodies raised against two major stress proteins, stress 70 and chaperonin 60, in diverse species

Journal Article · Mon Aug 01 00:00:00 EDT 1994 · Environmental Toxicology and Chemistry; (United States)

DOI:https://doi.org/10.1002/etc.5620130805· OSTI ID:7159161

Sanders, B M; Martin, L S; Nakagawa, P A ^[1]; Hunter, D A ^[2]; Miller, S ^[3]; Ullrich, S J ^[4]

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1. California State Univ., Long Beach, CA (United States)
2. Odense Univ. (Denmark). Biologisk Inst.
3. Dept. of Agriculture, Gainesville, FL (United States)
4. National Inst. of Health, Bethesda, MD (United States). National Cancer Inst.

Immunoblot analysis using several antibodies raised against two major families of stress proteins, stress 70 and chaperonin 60 (cpn60), which are highly conserved in mammals, was carried out in diverse species often used in environmental research, including molluscs, annelids, crustaceans, echinoderms, and fish. The study revealed surprisingly different patterns of antibody cross-reactivity among species. The monoclonal anti-stress 70 antibody (mAb) C92 was the least cross-reactive for all species tested. The mAbs anti-stress 70 N27, BRM-22, and 3a3 were more broadly cross-reactive, but their binding specifities to stress 70 isoforms in the diverse species tested did not correlate with one another or follow taxonomic lines. The polyclonal anti-stress 70 antibody reacted to proteins in the 70 to 74 kDa range in all fish examined and in most invertebrates. When a polyclonal antibody (pAb) raised against cpn60 from a moth was used as a probe, specific binding was observed with proteins in the 60 to 64 kDa range in all fish examined and in most invertebrates. However, the size and number of isoforms that reacted with the pAb were species specific. These data suggest that these two major stress protein families are less highly conserved in invertebrates and fish than in mammals. Therefore, to minimize misinterpretation when using antibodies in heterologous assays with species in which the stress response has not been well characterized, it is important to determine which isoforms of stress 70 react with a particular antibody and to take into account the differential regulation of each member of this multigene family.

• OSTI ID: 7159161
• Journal Information: Environmental Toxicology and Chemistry; (United States), Vol. 13:8; ISSN 0730-7268
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANNELIDS
BIOLOGICAL STRESS
CRUSTACEANS
ECHINODERMS
FISHES
MOLLUSCS
PROTEINS
BIOSYNTHESIS
TOXIC MATERIALS
BIOLOGICAL MARKERS
IMMUNOASSAY
ANIMALS
AQUATIC ORGANISMS
ARTHROPODS
BENTHOS
BIOASSAY
HAZARDOUS MATERIALS
INVERTEBRATES
MATERIALS
ORGANIC COMPOUNDS
SYNTHESIS
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology