| RRC ID |
36925
|
| Author |
Terasawa K, Rajapakshe AR, Podyma-Inoue KA, Mishima-Tsumagari C, Yanagishita M, Hara-Yokoyama M.
|
| Title |
Preferential recognition of isocitrate dehydrogenase by a rabbit monoclonal antibody (ab124797) against the C-terminal peptide of RANKL.
|
| Journal |
J Immunol Methods
|
| Abstract |
A rabbit monoclonal antibody (Abcam ab124797), with high affinity for a synthetic peptide corresponding to the C-terminal region of the receptor activator of nuclear factor (NF)-κB ligand (RANKL), specifically recognizes a 37 kDa protein by immunoblotting, in good agreement with the molecular mass of RANKL. However, our mass spectroscopy analysis revealed that the protein recognized by the antibody is the α-subunit of NAD(+)-dependent isocitrate dehydrogenase (ICDH), a key Krebs cycle enzyme in mitochondria. Consistently, immunocytochemical staining with the antibody revealed a network organization characteristic of mitochondria, which overlapped with staining by MitoTracker and was lost after the siRNA-mediated downregulation of ICDH. The C-terminal peptide of ICDH contains similar chemical characteristics to that of the RANKL peptide and interacts with the antibody, although the affinity is a hundred times weaker. The present study provides an example of the preferential recognition of a surrogate protein by a rabbit monoclonal antibody.
|
| Volume |
420
|
| Pages |
1-10
|
| Published |
2015-5-1
|
| DOI |
10.1016/j.jim.2015.03.006
|
| PII |
S0022-1759(15)00070-8
|
| PMID |
25771969
|
| MeSH |
Animals
Antibodies, Monoclonal / immunology*
Antibody Specificity*
Cross Reactions
Isocitrate Dehydrogenase / genetics
Isocitrate Dehydrogenase / immunology*
Mice
Protein Structure, Tertiary
RANK Ligand / genetics
RANK Ligand / immunology*
Rabbits
|
| IF |
1.901
|
| Times Cited |
0
|
|
WOS Category
|
BIOCHEMICAL RESEARCH METHODS
IMMUNOLOGY
|
| Resource |
| Human and Animal Cells |
MC3T3-E1(RCB1126)
HeLa |
