Glucagon Serum Assay Kit

Built for specificity, optimized for sensitivity, designed for ease of use.

Cisbio's glucagon assay offers superior specificity and sensitivity for a reliable quantification of glucagon in serum and plasma samples. Built on a truly homogeneous protocol, the assay does not require any washing or complex extraction steps and means significant time savings compared to Enzyme-linked immune-sorbent assay (ELISA) methods. Simply add the detection reagents, incubate, and read.

Assay principle

Insulin is measured using a sandwich immunoassay of two monoclonal antibodies, one labeled with Cryptate (Donor) and the other with d2 (Acceptor). The intensity of the FRET signal obtained is proportional to the concentration of glucagon in the sample.

Designed for ease of use, the assay does not require lengthy extraction procedures or sample pre-treatment, as may be the case for other technologies. Simply add the detection reagents and read. The two detection reagents may be pre-mixed for a single dispensing step.

Built for specificity, this assay is compatible with serum and plasma samples. 

Assay characteristics

Specificity

Cisbio has designed and optimized an assay with very high specificity and sensitivity for the quantification of glucagon in cell supernatant or plasma. Our assay has an extremely low cross-reactivity for oxyntomodulin, glicentin, and other glucagon related products such as miniglucagon, fragment 1-18, GLP-1, GLP-2 and GRPP, providing superior specificity for results you can trust.

Peptide Specificity ( % of recognition )
Glucagon 100%
Oxyntomodulin <0.07%
Glicentin <0.07%
Glucagon fragment 1-18 <1.81%
Glucagon fragment 19-29 <0.03%
GLP-1 (7-36) amide <0.06%
GLP-1 (7-37) <0.11%
GLP-2 <0.3%
GRPP (Glicentin-Related Pancreatic Peptide) <0.01%

Glucagon kit specificity: Cross-reactivity assessment on closely related peptides reveals an exceptional specificity profile.

Sensitivity

Typical calibration curves obtained by the dilution of glucagon standard in cell-culture media conditions for 20 µL final, using a white 384-well small volume plate, are presented below. The detection limit was 12 pg/mL, as measured on a PHERAstar FS reader (flash lamp excitation) .

  In diluent #6
Analytical sensitivity (= dose of mean zero +2 SD) ~12 pg/mL
Assay range 15.6 to 2,000 pg/mL
Incubation time Overnight at room temperature

 

Linearity

Dilution linearity in cell culture media was assessed, with recovery results between 100 % and 117%.

Serial dilutions of a human serum sample in diluent #6:

Human serum sample [Glucagon] measured (pg/mL) [Glucagon] expected (pg/mL) % of expected concentration
Undiluted 1666.7 - -
1:2 846.3 833.3 101.6 %
1:4 420.7 416.7 101.0 %
1:8 221.7 208.3 106.4 %
1:16 121.8 104.2 116.9 %

Human serum samples were collected and serially diluted in diluent #6. The results are expressed as percent observed from expected.

Intra and Inter assay variability

Intra-assay and inter-assay variability in cell culture media conditions were assessed. Typical CVs of less than 4 % - for intra - and less than or around 10% - for inter -  were obtained, supporting claims regarding the features of a robust and reproducible assay.

Intra-Assay - n = 24

CV

Standard 2 (= 31.25 pg/mL)

1.8%

Standard 6 (= 500 pg/mL)

1.9%

Standard 8 (= 2,000 pg/mL)

3.6%

Inter-Assay - n = 6

CV

Standard 2 (= 31.25 pg/mL)

11.2%

Standard 6 (= 500 pg/mL)

6.6%

Standard 8 (= 2,000 pg/mL)

6.4%

 

Assay validation

Normal and pathological levels of glucagon

Glucagon concentrations for 16 pathological and normal EDTA-plasma samples were determined using the HTRF Serum Glucagon kit reagents. The results obtained show the good discrimination of the glucagon assay,  clearly differentiating between normal and pathological levels of glucagon.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Glucagon Serum kit - 5 x 200 tests62SGLPEB
Glucagon Serum kit - 200 tests62SGLPEF
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Companion products

DescriptionCat. noProduct insertMSDS
Glucagon kit standard62GLCCDA
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