Development, evaluation, and application of a highly sensitive microtiter plate ELISA for human Bcl10 protein.
Bcl10 (B-cell CLL/lymphoma 10) is a 233 amino acid CARD (caspase recruitment domain)-containing cellular protein, increasingly recognized as a mediator of NFkappaB activation in non-immune, as well as immune cells. Due to the importance of Bcl10 in diverse cell types, we developed a solid-phase, enzyme-linked immunosorbent (ELISA) assay to precisely measure Bcl10 in small volume cell lysates, using recombinant Bcl10 to standardize the assay. Standard curve measures Bcl10 from 0.25 ng/mL to 16 ng/mL, with very low intra- and inter-assay variation. Sample dilution and exogenous Bcl10 recovery experiments, comparisons with Western blot, and linear response to increasing doses of known Bcl10 activators confirm the specificity and precision of the ELISA.
Bhattacharyya
Sumit
S
Department of Medicine, University of Illinois at Chicago and Jesse Brown VA Medical Center, Chicago, IL, 60612 USA.
Pant
Nitika
N
Dudeja
Pradeep K
PK
Tobacman
Joanne K
JK
D048868:Adaptor Proteins, Signal Transducing / Q000032:analysis*; D000074018:B-Cell CLL-Lymphoma 10 Protein; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D008832:Microchemistry / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D048868:Adaptor Proteins, Signal Transducing; D000074018:B-Cell CLL-Lymphoma 10 Protein; C118591:BCL10 protein, human