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Big Endothelin-1 ELISA

Cat. No.: BI-20082H

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Therapeutic Areas
BigET-1 standard curve
endostatin_1
gitEt-graph
Assay Characteristics
  • Category number

    BI-20082H

  • Method

    Sandwich ELISA, HRP/TMB, 12×8-well detachable strips

  • Sample type

    Serum, EDTA plasma, citrate plasma, heparin plasma

  • Sample volume

    50 µl / well

  • Assay time

    4 h / 1 h / 30 min

  • Sensitivity

    0.02 pmol/l (= 0.086 pg/ml)

  • Standard range

    0 – 3 pmol/l (= 0 – 12.85 pg/ml)

  • Conversion factor

    1 pg/ml = 0.2335 pmol/l (MW: 4.283 kDa)

  • Specificity

    Endogenous and recombinant Big Endothelin-1 (Big ET-1).

  • Precision

    In-between-run (n=5): ≤ 4 % CV

    Within-run (n=5): ≤ 5 % CV

  • Cross-reactivity

    Human ET1/2/3 (1-21): <1%, human ET2 (1-37): <1%, human ET1/2 (1-38): <1%, human BigET1/2 (22-38) : <1%, human BigET2 (22-37) : <1%,

    porcine BigET (1-39): 21%, rat BigET1 (1-39): 10%, Sarafotoxin: <1%

  • Use

    CE marked – for IVD use in the EU

  • Validation Data

    See validation data tab for: precision, accuracy, diltuion linearity, values for healthy donors, etc.

Product details

Product Overview

The Big Endothelin-1 immunoassay is a 5.5 hour, 96-well sandwich ELISA for the quantitative determination of Big Endothelin-1 in serum and plasma. The assay employs human serum-based standards to ensure the measurement of biologically reliable data.

Principle of the Assay

The Big Endothelin-1 ELISA kit is a sandwich enzyme immunoassay for the quantitative determination of Big Endothelin-1 in human serum and plasma samples.

The figure below explains the principle of the Big Endothelin-1 sandwich ELISA:

 

In a first step, standard/control/sample and detection antibody (monoclonal mouse anti- human Big Endothelin-1 antibody) are pipetted into the wells of the microtiter strips, which are pre-coated with polyclonal sheep anti-human Big Endothelin-1 antibody. Big Endothelin-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody.  After a washing step, which removes all non-specifically bound and unbound material, the conjugate (streptavidin-HRP) is pipetted into the wells and reacts with the detection antibody. After another washing step, the substrate (tetramethylbenzidine, TMB) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of Big Endothelin-1. This color change is detectable with a standard microtiter plate reader. The concentration of Big Endothelin-1 in the sample is determined directly from the dose response curve.

The antibodies utilized in the Big Endothelin-1 ELISA (BI-20082H) are as follows:

Capture Antibody: polyclonal sheep-anti human Big Endothelin-1 
Detection Antibody: monoclonal mouse anti-human Big Endothelin-1

Typical Standard Curve

The figure below shows a typical standard curve for the Big Endothelin-1 ELISA. The immunoassay is calibrated against recombinant Big Endothelin-1 peptide:



ELISA Kit Components   

 

Contents

Description

Quantity

PLATE

Polyclonal sheep anti-human Big Endothelin-1 antibody pre-coated microtiter strips in strip holder, packed in aluminum bag with desiccant

12 x 8 tests

WASHBUF

Wash buffer concentrate 20x, natural cap

1 x 50 ml

STD

Standards 1-6, (0; 0.10; 0.20; 0.40; 1; 3 pmol/l), synthetic human Big Endothelin-1 in human serum, white caps, lyophilized

6 vials

CTRL

Control, yellow cap, lyophilized, exact concentration see label

1 vial

AB

Monoclonal mouse anti-human Big Endothelin-1 antibody, biotin labeled, red dye, green cap, ready to use

1 x 18 ml

CONJ

Conjugate, (streptavidin-HRP), amber cap, ready to use

1 x 22 ml

SUB

Substrate, (TMB solution), blue cap, ready to use

1 x 22 ml

STOP

Stop solution, white cap, ready to use

1 x 7 ml

Storage instructions: All reagents of the Big Endothelin-1 ELISA kit are stable at 4°C until the expiry date stated on the label of each reagent.

Instructions for use

Sample Collection & Storage

Serum, EDTA plasma, heparin plasma, citrate plasma, cell culture supernatant and urine are suitable for use in this assay. Do not change sample type during studies. We recommend duplicate measurements for all samples, standards and controls. The sample collection and storage conditions listed are intended as general guidelines.
 

Serum & Plasma

Collect venous blood samples in standardized serum separator tubes (SST) or standardized blood collection tubes using EDTA, heparin or citrate as an anticoagulant. For serum samples, allow samples to clot for 30 minutes at room temperature. Perform separation by centrifugation according to the tube manufacturer’s instructions for use. Assay the acquired samples immediately or aliquot and store at -25°C or lower. Lipemic or haemolyzed samples may give erroneous results. Do not freeze-thaw samples more than four times.

 

Reagent Preparation

Wash Buffer

1.

Bring the WASHBUF concentrate to room temperature. Crystals in the buffer concentrate will dissolve at room temperature (18-26°C).

2.

Dilute the WASHBUF concentrate 1:20, e.g. 50 ml WASHBUF + 950 ml distilled or deionized water. Only use diluted WASHBUF when performing the assay.

The diluted WASHBUF is stable up to one month at 4°C (2-8°C).
 

Standards & Controls for Serum, Plasma and Urine Measurements

1.

Pipette 250 µl of distilled or deionized water into each standard (STD) and control (CTRL) vial. The exact concentration is printed on the label of each vial.

2.

Leave at room temperature (18-26°C) for 15 min. Vortex gently.

Reconstituted STDs and CTRL are stable at -25°C or lower until expiry date stated on the label. Avoid freeze-thaw cycles!
 

Sample Preparation

Bring samples to room temperature and mix samples gently to ensure the samples are homogenous. We recommend duplicate measurements for all samples.

Samples for which the OD value exceeds the highest point of the standard range can be diluted with STD 1 or Big Endothelin-1 negative human serum.

Assay Protocol

 

Read the entire protocol before beginning the assay.

1.

Bring reagents and samples to room temperature (18-24°C). 

2.

Mark positions for STD/CTRL/SAMPLE (standard/control/sample) on the protocol sheet.

3.

Take microtiter strips out of the aluminum bag. Store unused strips with desiccant at 4°C in the aluminum bag. Strips are stable until expiry date stated on the label.

4.

Add 50 µl STD/CTRL/SAMPLE (standard/control/sample) in duplicates into the respective wells.

5.

Add 150 µl AB (biotinylated anti-Big Endothelin-1 antibody, green cap) into each well. Swirl gently.

4.

Cover the plate tightly, swirl gently and incubate for 4 hours at room temperature (18-24°C) in the dark.

5.

Aspirate and wash wells 5 x with 300 µl diluted WASHBUF. After the final wash, remove the remaining WASHBUF by strongly tapping plate against a paper towel.

6.

Add 200 µl CONJ (conjugate, amber cap) into each well.

7.

Cover tightly and incubate for 1 hour at room temperature in the dark.

8.

Aspirate and wash wells 5 x with 300 µl diluted WASHBUF. After the final wash, remove remaining WASHBUF by strongly tapping plate against a paper towel.

9.

Add 200 µl SUB (substrate, blue cap) into each well.

10.

Incubate for 30 min at room temperature in the dark.

11.

Add 50 µl STOP (stop solution, white cap) into each well.

12.

Measure absorbance immediately at 450 nm with reference 630 nm, if available.

Calculation of Results

Read the optical density (OD) of all wells on a plate reader using 450 nm wavelength (reference wavelength 630 nm). If the OD of the highest STD is outside the measuring range of photometer the plate can be re-measured at 405 nm (correction wavelength 630 nm). Construct a standard curve from the absorbance read-outs of the standards using commercially available software capable of generating a four-parameter logistic (4-PL) fit. Alternatively, plot the standards’ concentration on the x-axis against the mean absorbance for each standard on the y-axis and draw a best fit curve through the points on the graph. Curve fitting algorithms other than 4-PL have not been validated and will need to be evaluated by the user.
Obtain sample concentrations from the standard curve. If required, pmol/l can be converted into pg/ml by applying a conversion factor (1 pg/ml =0.2335 pmol/l (MW: 4.283 kDa)). Respective dilution factors have to be considered when calculating the final concentration of the sample.

The quality control protocol supplied with the kit shows the results of the final release QC for each kit at production date. Data for OD obtained by customers may differ due to various influences and/or due to the normal decrease of signal intensity during shelf life. However, this does not affect validity of results as long as an OD of 1.00 or higher is obtained for the standard with the highest concentration and the control value is in range (target range see label).

Background & Therapeutic Areas

INFORMATION ON THE ANALYTE

Big Endothelin-1 Protein

Big Endothelin-1 (BigET-1) is a peptide of 38 amino acids and is the precursor of Endothelin-1 (ET-1), consisting of amino acids 1-21 (http://www.uniprot.org/uniprot/P05305). ET-1 is a potent vasoconstrictor and is produced by vascular endothelial cells. Accordingly, it has a wide tissue distribution (http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Hs.511899). The cleavage of BigET-1 by Endothelin-converting enzyme (ECE) leads to Endothelin (ET) and to a C‑terminal fragment.

The half-life of ET-1 (1-21) in plasma is less than one minute, which may preclude its use as a diagnostic marker, whereas clearance of the precursor peptide BigET-1 is much slower. As Big-Endothelin-1 is produced at amounts equimolar to ET-1 and has higher plasma stability, it is more suitable for diagnostics and monitoring of heart failure (Gergei et al., 2018). Hence, Big ET-1 has the same physiological role as ET-1 while beeing a more reliable marker.

Molecular Weight

4.283 kDa

Cellular localisation

Secreted

Post-translational modifications

Disulphide bonds, post-translational processing

Sequence similarities

Endothelin family

Alternative Names

For endothelin gene: Endothelin 1, Preproendothelin-1, Endothelin-1, PPET1, ARCND3, HDLCQ7, ET1, QME

Entrez/NCBI ID

EDN1: https://www.ncbi.nlm.nih.gov/gene/1906

Genecards

EDN1: https://www.genecards.org/cgi-bin/carddisp.pl?gene=EDN1&keywords=edn1

OMIM

131240: http://omim.org/entry/131240

PDB

1T7H: http://www.rcsb.org/structure/1T7H ;
3RDY:http://www.rcsb.org/structure/3RDY;
3RDZ: http://www.rcsb.org/structure/3RDZ ;

Pfam

Endothelin (PF00322): https://pfam.xfam.org/family/endothelin

Protein Atlas

EDN1 - https://www.proteinatlas.org/ENSG00000078401-EDN1/cell

Uniprot ID

EDN1 (P05305) - http://www.uniprot.org/uniprot/P05305

Big Endothelin-1 Function

Preproendothelin, a 203-amino-acid (AA) peptide is encoded by the ET-1 gene. It is converted into Big Endothelin-1 (Big ET-1), containing 38 amino acids. Big-ET-1 is then converted by the endothelin-converting enzyme (ECE) into the 21 AA biologically active Endothelin-1 peptide (AA1-21). Endothelin-1 circulates in low concentrations and has a very short plasma half-life. Big-Endothelin-1, the precursor is a stable peptide with a plasma half-life of about 30 minutes and measurement of Big ET-1 reflects Endothelin levels.

ET-1 is secreted, as a paracrine hormone, mainly from endothelial cells towards the smooth muscle cell layer of the vessels, to produce immediate vasoconstriction. Endothelin-1 (ET-1) has in fact been described as the most potent vasoconstrictor substance identified to date, and overactivation or dysfunction of the ET system has been implicated in the development and progression of hypertension in both clinical and experimental studies (Gillis EE et al., 2016). Big endothelin-1 has been demonstrated to be correlated with inflammation, endothelial dysfunction and arterial stiffness (Zhou BY et al., 2017). Both Big Endothelin and Endothelin are strong independent predictors of survival in patients with congestive heart failure and identify a population with very high-risk mortality. The pathophysiological roles of endothelin’s in the development of cardiovascular disease have been reviewed (Ohkita et al., 2012). It has been suggested that ET-1 is a major factor in the development of chronic kidney disease (CKD) as it contributes to hypertension, proteinuria, and renal inflammation in CKD. Numerous studies have demonstrated that ET-1 directly stimulates inflammation both in the vasculature and in the kidney, and this occurs in the absence of hypertension (Speed and Pollack, 2012).

Endothelins act via activation of two receptor subtypes, ETA and ETB receptors. Endothelin antagonists are currently used clinically in the treatment for patients with pulmonary hypertension (Davenport AP et al., 2016, Houde M et al., 2016), and are considered to have further target diseases as heart failure, cardiac hypertrophy and other cardiac diseases, renal diseases, systemic hypertension, and cerebral vasospasm (Miyauchi T and S Sakai, 2018).

  • Cancer

    • Oral squamous cell carcinoma (Mankapure et al., 2015)

    • Endothelin therapeutics in cancer (Rosanò and Bagnato, 2016)

  • Cardiology-Cardiovascular diseases

    • Prognostic value in heart failure and acute myocardial infarction (Gergei et al., 2017; Jankowich et al., 2016; Olivier et al., 2017)

    • Cardiomyopathy (Matsa et al., 2014; Yilu Wang et al., 2017)

    • Coronary artery disease and atherosclerosis (Chen et al., 2015; Qing et al., 2015; Wang et al., 2018; Yao Wang et al., 2017; Zhou et al., 2017)

    • Pulmonary hypertension (Yoshibayashi et al., 1991)

    • Pumonary thromboendarterectomy (Langer et al., 2005)

    • Cardiovascular disease (Ohkita et al., 2012)

    • Cardiovascular events (Jankowicha and Choudhary, 2019)

  • Nephrology

    • Renal insufficiency (Fischer et al.,2017)

    • During and after graft rejection (Jeremy et al., 2011)

    • Kidney disease and hypertension (Speed and Pollack, 2012)

Validation Data

All Biomedica ELISAs are validated according to FDA/ICH/EMEA guidelines. For more information about our validation guidelines, please refer to our quality page and published validation guidelines and literature.

Validation literature

1. ICH Q2(R1) Validation of Analytical Procedures: Text and Methodology.
2. EMEA/CHMP/EWP/192217/2009 Guideline on bioanalytical method validation.
3. Bioanalytical Method Validation, Guidance for Industry, FDA, May 2018

Calibration

This immunoassay is calibrated against recombinant human Big Endothelin-1 (1-38).

Detection Limit & Sensitivity

To determine the sensitivity of the Big Endothelin-1 ELISA, experiments measuring the lower limit of detection (LOD) and the lower limit of quantification (LLOQ) were conducted.

The LOD, also called the detection limit, is the lowest point at which a signal can be distinguished above the background signal, i.e. the signal that is measured in the absence of Big Endothelin-1, with a confidence level of 99%. It is defined as the mean back calculated concentration of standard 1 (0 pmol/l of Big Endothelin-1) plus three times the standard deviation of the measurements.

The LLOQ, or sensitivity of an assay, is the lowest concentration at which an analyte can be accurately quantified. The criteria for accurate quantification at the LLOQ are an analyte recovery between 75 and 125% and a coefficient of variation (CV) of less than 25%. To determine the LLOQ, standard 2, i.e. the lowest standards containing Big Endothelin-1, is diluted, measured and its concentration back calculated. The lowest dilution, which meets both criteria, is reported as the LLOQ.

The following values were determined for the Big Endothelin-1 ELISA:

LOD

0.02 nmol/l

LLOQ

0.03 nmol/l

Precision

The precision of an ELISA is defined as its ability to measure the same concentration consistently within the same experiments carried out by one operator (within-run precision or repeatability) and across several experiments using the same samples but conducted by several operators at different locations using different ELISA lots (in-between-run precision or reproducibility).

Within-Run Precision

Within-run precision was tested by measuring two samples 5 times within one human Big ET-1 ELISA lot. The experiment was conducted by one operator.

ID

 

Within-Run Precision

n

Mean NT-proANP [pmol/l]

SD [pmol/l]

CV (%)

Sample 1

5

0.20

0.003

2

Sample 2

5

1.00

0.048

5

In-Between-Run Precision

In-between-run precision was assessed by measuring two samples ten times within three human BigET-1 ELISA kit lots by three different operators.

ID

 

In-Between Run Precision

n

Mean NT-proANP [pmol/l]

SD [pmol/l]

CV [%]

Sample 1

10

0.20

0.009

4

Sample 2

10

1.0

0.041

4

Accuracy

The accuracy of an ELISA is defined as the precision with which it can recover samples of known concentrations.

 

The recovery of the Big Endothelin-1 ELISA was measured by adding recombinant Big Endothelin-1 to human samples containing a known concentration endogenous Big Endothelin-1. The %recovery of the spiked concentration was calculated as the percentage of measured compared over the expected value. All our ELISAs are expected to have %recovery rates within 15% of the nominal value of the sample.

 

This table shows the summary of the recovery experiments in the Big Endothelin-1 ELISA in different sample matrices:

   

% Recovery

Sample Matrix

n

+ 1 pmol/l

+ 2 pmol/l

Mean

Range

Mean

Range

Serum

14

100

80 - 127

105

85 – 119

EDTA plasma

3

100

91 – 108

101

95 – 114

Citrate plasma

3

98

83 -111

105

96 - 114

Heparin plasma

3

97

92 – 108

102

96 – 109

Experiments:

Data showing recovery of recombinant Big Endothelin-1 in human serum samples:

   

Big Endothelin-1 [pmol/l]

% Recovery

Sample Matrix

ID

Reference

+ 1 pmol/l

+ 2 pmol/l

+ 1 pmol/l

+ 2 pmol/l

Serum

s1

0.65

1.6

2.7

97

102

Serum

s2

0.8

2.1

2.9

117

104

Serum

s3

0.02

1.3

2.4

127

119

Serum

s4

0.3

1.5

2.4

115

104

Serum

s5

0

1.1

2.5

110

125

Serum

s6

0

0.9

2.2

90

110

Serum

s7

0

1.0

2.3

100

115

Serum

s8

0

0.8

1.7

80

85

Serum

s9

0

0.8

1.9

80

95

Serum

s10

0

1.1

1.9

110

95

Serum

s11

0

1.0

2.1

100

105

Serum

s12

0.1

1.1

2.3

100

110

Serum

s13

0.06

0.9

1.8

85

87

Serum

s14

0.06

0.9

2.3

85

112
       

Mean

100

105
       

Min

80

85
       

Max

127

119

Data showing recovery of recombinant Big Endothelin-1 in human EDTA plasma samples:

   

Big Endothelin-1 [pmol/l]

% Recovery

Sample Matrix

ID

Reference

+ 1 pmol/l

+ 2 pmol/l

+ 1 pmol/l

+ 2 pmol/l

EDTA plasma

e1

0.11

1.2

2.4

108

114

EDTA plasma

e2

0.09

1.1

2.1

101

100

EDTA plasma

e3

0.1

1.0

2.0

91

95
       

Mean

100

101

Data showing recovery of recombinant Big Endothelin-1 in a human citrate plasma sample:

   

Big Endothelin-1 [pmol/l]

% Recovery

Sample Matrix

ID

Reference

+ 1 pmol/l

+ 2 pmol/l

+ 1 pmol/l

+ 2 pmol/l

Citrate plasma

c1

0.11

1.1

2.4

99

114

Citrate plasma

c2

0.08

1.2

2.2

111

106

Citrate plasma

c3

0.08

0.9

2.0

83

96
       

Mean

98

105

Data showing recovery of recombinant Big Endothelin-1 in a human heparin plasma sample:

   

Big Endothelin-1 [pmol/l]

% Recovery

Sample Matrix

ID

Reference

+ 1 pmol/l

+ 2 pmol/l

+ 1 pmol/l

+ 2 pmol/l

Heparin plasma

h1

0.11

1.2

2.3

108

109

Heparin plasma

h2

0.09

1.0

2.0

92

96

Heparin plasma

h3

0.08

1.0

2.1

93

101
       

Mean

97

102

Dilution Linearity & Parallelism

Tests of dilution linearity and parallelism ensure that both endogenous and recombinant samples containing Big Endothelin-1 behave in a dose dependent manner and are not affected by matrix effects. Dilution linearity assesses the accuracy of measurements in diluted human samples spiked with known concentrations of recombinant analyte. By contrast, parallelism refers to dilution linearity in human samples and provides evidence that endogenous analyte behaves same way as the recombinant one. For dilution linearity and parallelism are assessed for each sample type and are considered good if the results are within 20% of the expected concentration.

Dilution linearity was assessed by serially diluting human samples spiked with 2 pmol/l recombinant Big Endothelin-1 with standard 1 (human serum containing 0 pmol/l Big Endothelin-1).

The table below show the mean recovery and range of serially diluted recombinant Big Endothelin-1 in serum:

   

% Recovery of recombinant Big Endothelin-1 in diluted samples

Sample

Matrix

n

1+1

1+3

Mean

Range

Mean

Range

Serum

8

90

62- 105

96

71 - 126

Data showing dilution linearity of recombinant Big Endothelin-1 spiked into human blood samples (ref) containing endogenous Big Endothelin-1:

   

Big Endothelin-1 [pmol/l]

% Recovery

Sample Matrix

ID

Ref

1+1

1+3

1+1

1+3

Serum

s1

1.36

0.42

0.24

62

71

Serum

s2

0.24

0.13

0.08

105

126

Serum

s3

0.48

0.25

0.13

103

110

Serum

s4

1.46

0.64

0.33

87

92

Serum

s5

0.78

0.37

0.20

94

102

Serum

s6

1.43

0.56

0.31

79

87

Serum

s7

3.11

1.47

0.59

95

76

Serum

s8

2.67

1.28

0.69

96

104
       

Mean

90

96
       

Min

62

71
       

Max

105

126

Parallelism was assessed by serially diluting human samples containing endogenous Big Endothelin-1 with with standard 1 (human serum containing 0 pmol/l Big Endothelin-1).

The table below show the mean recovery and range of serially diluted endogenous Big Endothelin-1 in several sample matrices:

   

% Recovery of endogenous Big Endothelin-1 in diluted samples

Sample

Matrix

n

1+1

1+3

Mean

Range

Mean

Range

EDTA plasma

4

110

99 – 127

104

91 - 124

Data showing dilution linearity of endogenous Big Endothelin-1 in human EDTA plasma samples:

   

Big Endothelin-1 [pmol/l]

% Recovery

Sample Matrix

ID

Ref

1+1

1+3

1+1

1+3

EDTA plasma

e1

2.06

1.30

0.64

127

124

EDTA plasma

e2

2.58

1.38

0.63

107

97

EDTA plasma

e3

2.30

1.23

0.61

107

106

EDTA plasma

e4

2.64

1.31

0.60

99

91
       

Mean

110

104
       

Min

99

91
       

Max

127

124

Specificity

This assay recognizes endogenous (natural) and recombinant human Big Endothelin-1 (Big ET-1).

Cross Reactivity

The factors listed below were assayed for cross-reactivity:

  • Human ET1/2/3 (1-21): <1%
  • Human ET2 (1-37): <1%
  • Human ET1/2 (1-38): <1%
  • Porcine BigET (1-39): 21%
  • Human BigET1/2 (22-38): <1%
  • Human BigET2 (22-37): <1%
  • Rat BigET1 (1-39): 10% 
  • Sarafotoxin: <1%

 

Sample Stability

The stability of endogenous Big Endothelin-1 was tested by comparing Big Endothelin-1 measurements in samples that had undergone up to four freeze-thaw cycles.

For freeze-thaw experiments, samples were collected according to the supplier’s instruction using blood collection devices and stored at -80°C. Reference samples were freeze-thawed once. The mean recovery of sample concentration after four freeze-thaw cycles is 109%.

 

Big Endothelin-1 [pmol/l]

% Recovery after 4 freeze/thaw cycles

ID

1 x

2 x

3 x

4 x

s1

0.75

0.80

0.79

0.84

111 

s2

0.15

0.14

0.12

0.17

120

s3

0.15

0.13

0.14

0.16

103

s4

0.84

0.79

0.86

0.84

100
       

Mean

109

Samples can undergo at least up to 3 freeze-thaw cycles.
 

Sample Values

To provide expected values for circulating Big Endothelin-1, a panel of samples from apparently healthy donors as well as samples from patient cohorts with cardiovascular and renal diseases were tested.

A summary of the results is shown below:

   

Big Endothelin-1 [pmol/l]

Sample Matrix

n

Mean

Median

Minimum

Maximum

Apparently healthy

41

0.22

0.09

0.01

1.58

Cardio panel

30

0.8

0.6

0.2

2.1

Dialysis panel

36

1.1

0.9

0.4

3.0

It is recommended to establish the normal range for each laboratory.

NT-proANP Values in an unselected hospital panel

NT-proANP was measured in patients from an unselected hospital panel. No medical histories were available.

Sample Matrix

EDTA-plasma

n

40

Mean [nmol/l]

3.89

Median [nmol/l]

3.09

Min [nmol/l]

0.63

Max [nmol/l]

10.50

Matrix Comparision

To assess whether all tested matrices behave the same way in the Big Endothelin-1 ELISA, concentrations of Big Endothelin-1 were measured in serum, EDTA, heparin, and citrate plasma samples prepared from three apparently healthy donors. Each individual donated blood in all tested sample matrices.

A summary table of Big Endothelin-1 levels in various sample matrices is shown below:

 

Big Endothelin-1 [pmol/l]

  

Sample ID

Serum

EDTA plasma

Citrate plasma

Heparin plasma

% CV

#1

0.1

0.11

0.11

0.11

7

#2

0.06

0.09

0.09

0.08

17

#3

0.06

0.1

0.08

0.08

23

A figure of Big Endothelin-1 levels in various sample matrices is shown below:



0:21 / 3:06

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