Expression and localisation of stanniocalcin 1 in rat bladder, kidney and ovary.
Bony fish use the glycoprotein hormone stanniocalcin (STC) to counteract hypercalcaemia. This is achieved through dual mechanisms involving gill calcium uptake inhibition and stimulation of renal inorganic phosphate reabsorption. Human STC (hSTC-1) shows considerable homology with both rat and mouse STC (mSTC) and their mRNA is expressed in a wide range of tissues. In fish, STC is produced by endocrine glands known as the corpuscles of Stannius but in mammals the widespread expression is suggestive of a paracrine rather than an endocrine role. In order to determine the distribution and strucutral characteristics of hSTC-1, the recombinant protein was expressed in bacteria, purified by metal-ion affinity chromatography, and a study was made of the likely epitopes for raising an antibody. This novel hSTC-1 antibody was used to test the purification protocol. Since the role of mammalian STC is largely unknown, the specific distribution of STC needed to be addressed. To test the specificity of the antibody, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting was undertaken in homogenised rat bladder, ovary and kidney.
Worthington
R A
RA
Institute for Biomedical Research, Department of Anatomy and Histology, The University of Sydney, NSW, Australia.
Brown
L
L
Jellinek
D
D
Chang
A C
AC
Reddel
R R
RR
Hambly
B D
BD
Barden
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D006728:Hormones / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D006801:Humans; D007668:Kidney / Q000737:chemistry*; D051379:Mice; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D010053:Ovary / Q000737:chemistry*; D010446:Peptide Fragments / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000276:immunology; D001743:Urinary Bladder / Q000737:chemistry*
D006023:Glycoproteins; D006728:Hormones; D010446:Peptide Fragments; D011994:Recombinant Proteins; C051795:teleocalcin