Immuno-Diagnostics Antibodies & Antigens for Veterinary Cardiovascular Diseases Test

Immunoassay calibration curve for NT-proBNP

These immunoassays have a sensitivity of 22 pg/mL for measuring recombinant NT-proBNP. Figures 1 and 2 display the suggested pairing calibration curves. Conversion of 1NT-proBNP concentration: 1 pmol/L = 1÷10.545 pg/ml Please note that a variety of factors might affect how well an immunoassay test performs. These consist of the labeling procedure, the kind of label coupled to the detection antibody, and the detection platform. Thus, on our customers' immunoassay systems, other NT-proBNP antibody pairs with non-overlapping epitopes could perform better.

Figure 1-2. NT-proBNP immunoassay calibration curve

(left) Calibration curve for optimal paired detection
(right) Calibration curve and canine plasma sample dilution comparison

Reagent type: two-step sandwich immunofluorescence detection, streptavidin-coated microplate
Capture antibody: 200ng/well, conjugated biotin
Capture antibody: 200ng/well, labeled europium chelate
Antigen: Canine recombinant NT-proBNP (Cat.#GMP-CAN-NT-proBNP-Ag01)
Sample volume: 50μl
Incubation time: 40 minutes at room temperature

Sensitivity detection of NT-proBNP

These immunoassays have a sensitivity of 22 pg/ml 1 for measuring recombinant NT-proBNP. Figures 1 and 2 display the suggested pairing calibration curves. Conversion of 1 NT-proBNP concentration: 1 pmol/L = 1÷10.545 pg/ml Please be aware that a variety of factors might affect how well an immunoassay test performs. These consist of the labeling procedure, the kind of label coupled to the detection antibody, and the detection platform. Thus, on our customers' immunoassay systems, other NT-proBNP antibody pairs with non-overlapping epitopes could perform better.

Figure 3. NT-proBNP concentration in EDTA plasma samples from healthy dogs and dogs with heart disease.

Reagent type: two-step sandwich immunofluorescence detection, streptavidin-coated microplate
Capture antibody: GMP-CAN-NT-proBNP-Ab01, 1μg/well
Capture antibody: GMP-CAN-NT-proBNP-Ab02, 200ng/well, labeled europium chelate
Calibrator: Canine recombinant NT-proBNP (Cat.# GMP-CAN-NT-proBNP-Ag01)
Sample volume: 50μl
Incubation time: 40 minutes at room temperature

Stability Test of NT-proBNP

Our antibodies were tested for their ability to detect endogenous canine NT-proBNP during sample storage. EDTA plasma from dogs with heart disease was incubated at two temperatures: +4°C and +20°C. At +4°C, NT-proBNP remains stable for at least 72 hours (95%–105% of initial immunoreactivity, Figure 4A). Simultaneously, at +20°C, about 98% of the initial immunoreactivity is detectable after 24 hours (Figure 4B). Using our recommended antibody pairings, plasma can be stored at +4°C for at least 72 hours with minimal loss of immunoreactivity. However, at room temperature, there is a slight weakening of the signal during the first 24 hours. Note that individual plasma or serum samples may vary.

Figure 4-5. Stability of endogenous canine NT-proBNP in EDTA plasma

(left) Immunoreactivity of NT-proBNP in plasma samples incubated for 24, 48 and 72 hours at +4°C.
(right) Immunoreactivity of NT-proBNP in plasma samples incubated for 4, 8, 24 and 48 hours at +20°C.

Collect protease inhibitor-free EDTA plasma; samples are centrifuged, separated, and stored at -70°C before use. EDTA plasma is stored at +4°C or +20°C with the addition of 0.1% NaN3 to prevent bacterial growth. After incubation, samples were stored at -70°C before measurement. The NT-proBNP concentration in plasma was 9 ng/ml (determined by GMP-CAN-NT-proBNP-Ab01 and GMP-CAN-NT-proBNP-Ab02 immunoassays). Detection using GMP-CAN-NT-proBNP-Ab01 and GMP-CAN-NT-proBNP-Ab05 as capture antibodies and GMP-CAN-NT-proBNP-Ab06 and GMP-CAN-NT-proBNP-Ab02 as detection antibodies respectively The system results are depicted in Figure 3.