DRG International EIA-2395 Leptin Sandwich Elisa Instructions

DRG International EIA-2395 Leptin Sandwich

Please use only the valid version of the Instructions for Use provided with the kit.

Introduced modifications

The following changes have been made in comparison to the previous version:

Addition of French version

INTENDED USE

The DRG Leptin Sandwich ELISA is a manual enzyme immunoassay for the quantitative measurement of leptin in human serum or Li-heparin plasma.
For in vitro diagnostic use. For laboratory professional use.
The device is intended to be used as an aid to the diagnosis of adiposity, leptin resistance, and congenital leptin deficiency.

Scientific Validity

Leptin is produced primarily in the adipocytes of white adipose tissue and circulates in blood in free form and bound to proteins (1). In mammals, leptin is pleiotropic, regulating a multitude of physiological processes. Leptin reduces appetite and food intake, and inhibits hepatic glucose production, fatty acid synthesis and the expression of resistin. In contrast, Leptin increases energy expenditure by inducing oxidation of fatty acids in liver and muscle. Moreover, Leptin stimulates  insulin secretion and glucose uptake as well as secretion of inflammatory cytokines (2,3). Leptin serves as a lipo static signal and conveys critical information regarding metabolic state to the brain by stimulating anorexic  proopiomelanocortin/cocaine and amphetamine-related transcript neurons and inhibiting orexigenic neuropeptide Y/ agouti-related protein neurons (4,5). The actions of Leptin are opposed by the hormone ghrelin. Both hormones act on receptors in the arcuate nucleus of the hypothalamus to regulate appetite to achieve energy homeostasis (6).
Although leptin reduces appetite as a circulating signal, obese individuals generally exhibit a higher circulating concentration of leptin than normal weight individuals due to their higher percentage body fat (7). These people show resistance to leptin, similar to resistance of insulin in type 2 diabetes, with the elevated levels failing to control hunger and modulate their weight. In obesity, a decreased sensitivity to leptin occurs, resulting in an inability to detect satiety despite high energy stores (8). Although regulation of fat stores is deemed to be the primary function of leptin, it also plays a role in other physiological processes, as evidenced by its multiple sites of synthesis other than fat cells, and the multiple cell types beside hypothalamic cells that have leptin receptors.
Leptin-deficient pathologies are typically accompanied by hyperphagia and obesity (9,10). Extreme obesity can be observed with mutations in the leptin receptor. The anorexigenic properties of leptin have been well characterized in the context of leptin-deficient humans, resulting in the reduction of food intake and body mass (11,12).
In conclusion, Leptin can be measured for the differential diagnosis of obesity with leptin resistance.

PRINCIPLE OF THE TEST

The DRG Leptin Sandwich ELISA is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle.
The microtiter wells are coated with a monoclonal antibody directed towards a unique antigenic site of the leptin molecule.
During the first incubation, leptin in the added sample binds to the immobilized antibody.
Unbound sample material is washed off.
In a second incubation added antiserum, which contains biotinylated anti- leptin antibody, forms a sandwich complex with the bound leptin on the microtiter wells.
Afterwards, the unbound antiserum is washed off and streptavidin peroxidase complex (Enzyme Complex) is added for detection of the biotin molecules.
After a washing step to remove all unbound substances, the solid phase is incubated with the substrate solution. The colorimetric reaction is stopped by addition of stop solution, and optical density (OD) of the resulting yellow product is measured. The intensity of color is proportional to the concentration of the analyte in the sample.
A standard curve is constructed by plotting OD values against concentrations of standards, and concentrations of unknown samples are determined using this standard curve.

WARNINGS AND PRECAUTIONS

• This kit is for in vitro diagnostic use only. For laboratory professional use only.
• Before starting the assay, read the instructions for use completely and carefully. Use the valid version of instructions for use provided with the kit. Be sure that everything is understood.
• Do not mix or use components from kits with different lot numbers. It is advised not to exchange wells of different plates even of the same lot. The kits may have been shipped or stored under different conditions and the binding characteristics of the plates may result slightly different.
• Do not use reagents beyond expiry date as shown on the kit labels.
• Do not reuse microtiter wells.
• Reagents of other manufacturers must not be used together with the reagents of this test kit.
• All reagents in this kit are clear liquids, substrate solution is clear and colorless. Changes in its appearance may affect the performance of the test. In that case, contact DRG.
• Microbial contamination of reagents or samples may give false results.
• Allow the reagents to reach room temperature (20 °C to 26 °C) before starting the test. Temperature will affect the optical density readings of the assay.
• All indicated volumes must be performed according to the protocol. Optimal test results are only obtained when using calibrated pipettes and microtiter plate readers.
• Use reservoirs only for single reagents. This especially applies to the substrate reservoirs. Using a reservoir for dispensing a substrate solution that had previously been used for the conjugate solution may turn solution coloured.
  Do not pour reagents back into original vials as reagent contamination may occur.

General precautions

• Follow good laboratory practice and safety guidelines.
• Never pipet by mouth and avoid contact of reagents and samples with skin and mucous membranes.
• Do not smoke, eat, drink, or apply cosmetics in areas where specimens or kit reagents are handled.
• Wear lab coats and disposable latex gloves when handling samples and reagents and safety glasses where necessary.

Biohazard information

• All reagents of this test kit which contain human serum or plasma have been tested and confirmed negative for HIV I/II, HBsAg and HCV by FDA approved procedures. However, no known test method can offer total assurance that no infectious agent is present.
• The device contains material of animal origin, which is certified apparently free of infectious or contagious diseases and injurious parasites.
• Bovine components originate from countries where BSE has not been reported.
• All materials and samples of human or animal origin must be handled as if capable of transmitting infectious diseases.
• Handling must be done in accordance with the procedures defined by appropriate national biohazard and safety guideline or regulation. Waste must be discarded according to local rules and regulations.

Information to chemical hazard and hazard classification

• Some reagents contain Pro Clin 300®, BND or MIT as preservative in non-declarable concentrations. Nevertheless, in case of contact with eyes or skin, flush immediately with water.
• Substrate Solution contains an ingredient in non-declarable concentrations which causes serious eye irritation. In case of possible contact with eyes, rinse immediately carefully and thoroughly with eye wash or water. After contact with skin, wash with plenty of water. Take-off contaminated clothing and wash it before reuse.
• Avoid contact with Stop Solution containing 0.5 M H2SO4. It may cause skin irritation and burns.
• Chemicals and prepared or used reagents must be treated as hazardous waste according to the national safety guideline or regulation.
• This product does not contain substances which have carcinogenic, mutagenic or toxic for reproduction (CMR) properties.

All reagents of this test kit do NOT contain hazardous substances in concentrations to be declared, a classification and labelling is not required.

For detailed information please refer to the Safety Data Sheet, which is available upon request directly from DRG.

MATERIALS

Materials provided with the kit

1. Microtiter wells, 12 x 8 strips, 96 wells (break apart); Wells coated with anti-leptin antibody (monoclonal).

2. Standard (Standard 0 – 5), 6 vials, 0.5 mL each, lyophilized; Concentrations: 0 – 2 – 5 – 25 – 50 – 100 ng/mL The standards are calibrated against the following reference material: WHO International Standard Leptin, human; NIBSC Code 97/594 See “Reagent Preparation“.
   Contain non-mercury preservative.

3. Control Low & High, 2 vials, 0.5 mL each, lyophilized; For control values and ranges please refer to vial label or Certificate of Analysis.
   See “Reagent Preparation“.
   Contain non-mercury preservative. Assay Buffer, 1 vial, 11 mL, ready to use; Contains non-mercury preservative.

4. Antiserum, 1 vial, 11 mL, ready to use, monoclonal biotinylated anti-Leptin antibody; Contain non-mercury preservative.

5. Enzyme Complex, 1 vial, 11 mL, ready to use; Streptavidin conjugated to horseradish peroxidase Contains non-mercury preservative.

6. Substrate Solution, 1 vial, 14 mL, ready to use; Contains 3,3‘,5,5‘-tetramethylbenzidine (TMB). Keep away from direct sun light.

7. Stop Solution, 1 vial, 14 mL, ready to use; Contains 0.5 M H2SO4, Avoid contact with the stop solution. It may cause skin irritations and burns.

8. Wash Solution, 1 vial, 30 mL (40X concentrated); See “Reagent Preparation“.

9. Instructions for Use

10. Certificate of Analysis (CoA)

Materials required but not provided

• A calibrated microtiter plate reader (450 nm, with reference wavelength at 620 nm to 630 nm)
• Calibrated variable precision micropipettes
• Manual or automatic equipment for rinsing microtiter plate wells
• Absorbent paper
• Distilled water
• Timer
• Graph paper or software for data reduction

Storage and Stability of the Kit

When stored at 2 °C to 8 °C unopened reagents will retain reactivity until expiration date.
Do not use reagents beyond this date.
Opened reagents must be stored at 2 °C to 8 °C.
Under the described storage conditions, opened kits retain their reactivity for 8 weeks.
The microtiter plate contains snap-off strips. Do not open the pouch of the wells until it reaches room temperature.
Unused wells must be stored at 2 °C to 8 °C in the sealed foil pouch including the desiccant and used in the frame provided. Once the foil bag has been opened, care must be taken to close it tightly again.

Reagent Preparation

Bring all reagents and required number of strips to room temperature (20 °C to 26 °C) prior to use.

Standards

Reconstitute the lyophilized contents of each standard vial with 0.5 mL distilled water and let stand for at least 10 minutes at room temperature. Mix several times before use.

Note: The reconstituted standards are stable for 6 weeks at 2 °C to 8 °C.
For longer storage the reconstituted standards must be aliquoted and stored frozen at -20 °C.

Controls

Reconstitute the lyophilized content of each vial with 0.5 mL distilled water and let stand for at least 10 minutes at room temperature. Mix the control several times before use.

Note: The reconstituted controls are stable for 6 weeks at 2 °C to 8 °C.
For longer storage the reconstituted controls must be aliquoted and stored frozen at -20 °C.

Wash Solution

Add distilled water to the 40X concentrated Wash Solution.
Dilute 30 mL of concentrated Wash Solution with 1170 mL distilled water to a final volume of 1200 ml.
The diluted Wash Solution is stable for 1 week at room temperature.

Disposal of the Kit

The disposal of the kit and all used materials/reagents must be performed according to the national regulations. Special information for this product is given in the Safety Data Sheet, section 13.

Damaged Test Kits

In case of any damage to the test kit or components, DRG must be informed in writing, at the latest one week after receiving the kit. Damaged single components must not be used for a test run. They have to be stored until a final solution has been found. After this, they must be disposed of according to the official regulations.

SAMPLE COLLECTION, STORAGE AND PREPARATION

The following sample material can be used in this test:
Human serum or lithium heparin plasma

Samples containing sodium azide should not be used in the assay.
In general, it should be avoided to use hemolytic, icteric, or lipaemic samples. For further information refer to chapter “Interfering Substances”.

Sample Collection

Serum: Collect blood by venipuncture (e.g. Sarstedt Monovette for serum), allow to clot, and separate serum by centrifugation at room temperature. Do not centrifuge before complete clotting has occurred. Patients receiving anticoagulant therapy may require increased clotting time.

Plasma: Whole blood should be collected into centrifuge tubes containing anticoagulant (e.g. Sarstedt Monovette with the appropriate plasma preparation) and centrifuged immediately after collection.

Samples Storage

Samples must be capped and can be stored for up to 2 months at 2 °C to 8 °C prior to performing the assay.
Samples stored for a longer time (up to 24 months) must be frozen only once at -20 °C prior to assay. Thawed samples must be inverted several times prior to testing.

Sample Dilution

If in an initial assay, a sample is found to contain more analyte than the highest standard, the sample can be diluted with Standard 0 and re-assayed as described in “Assay Procedure”.
For the calculation of the concentrations this dilution factor must be considered.

Example:

a) dilution 1: 10: 10 µL sample + 90 µL Standard 0; Mix thoroughly.
b) dilution 1 :100: 10 µL dilution a) 1:10 + 90 µL Standard 0; Mix thoroughly.

ASSAY PROCEDURE

Procedural Notes

• All reagents and samples must be allowed to come to room temperature (20 °C to 26 °C) before use.

• All reagents must be mixed without foaming.

• Do not interchange caps of reagent vials to avoid cross-contamination.

• Use new disposal plastic pipette tips for each standard, control, or sample in order to avoid carry-over.

• Mix the contents of the microtiter plate wells thoroughly to ensure good test results.

• Do not let wells dry during assay; add reagents immediately after completing the rinsing steps.

• Once the test has been started, all steps must be completed without interruption and in the same sequence for each step.

• The enzymatic reaction is linearly proportional to time and temperature.

• Optical density is a function of the incubation time and temperature. Respect the incubations times and temperatures as given in chapter “Test Procedure”.

• Before starting the assay, it is recommended that all reagents are ready, caps removed, all needed wells secured in holder, etc. This will ensure equal elapsed time for each pipetting step without interruption.

• Important note to wash procedure:
  Washing is critical. Improperly washed wells will give erroneous results. The sensitivity and precision of this assay is markedly influenced by the correct performance of the washing procedure!

• Test performance using fully automated analysis devices:
  Automated test performance using fully automated, open-system analysis devices is possible. However, the combination must be validated by the user.

Test Procedure

Each run must include a standard curve.
The controls serve as internal controls for the reliability of the test procedure. They must be assayed with each test run.
The given test procedure describes manual processing.

1. Secure the desired number of Microtiter wells in the frame holder.

2. Pipette 15 µL of each Standard, Control and sample with new disposable tips into appropriate wells.

3. Add 100 µL Assay Buffer into each well.
   Thoroughly mix for 10 seconds. It is important to have a complete mixing in this step.

4. Incubate for 120 minutes at room temperature.

5. Wash the wells as follows:
   If the wash step is performed manually:
   Briskly shake out the contents of the wells.
   Rinse the wells 3 times with 300 µL diluted Wash Solution per well.
   If an automated plate washer is used:
   Rinse the wells 3 times with 400 µL diluted Wash Solution per well.
   At the end of the washing step, always strike the wells sharply on absorbent paper to remove residual droplets.

6. Dispense 100 µL Antiserum to each well.

7. Incubate for 30 minutes at room temperature.

8. Wash as described in step 5.

9. Dispense 100 µL Enzyme Complex into each well.

10. Incubate for 30 minutes at room temperature.

11. Wash as described in step 5.

12. Add 100 µL of Substrate Solution to each well.

13. Incubate for 15 minutes at room temperature.

14. Stop the enzymatic reaction by adding 50 µL of Stop Solution to each well.

15. Measure the optical density of the solution in each well at 450 nm (reading) and at 620 nm to 630 nm (background subtraction, recommended) with a microtiter plate reader.
    It is recommended that the wells be read within 10 minutes after adding the Stop Solution.

Calculation of Results

1. Calculate the average optical density (OD) values for each set of standards, controls and patient samples.
2. Using linear graph paper, construct a standard curve by plotting the (mean) OD obtained from each standard against its concentration with OD value on the vertical (Y) axis and concentration on the horizontal (X) axis.
3. Determine the corresponding concentration from the standard curve by using the (mean) OD value for each sample.
4. Automated method: The results in the Instructions for Use have been calculated automatically using a 4-Parameter curve fit. (4-Parameter Rod bard or 4-Parameter Marquardt are the preferred methods.) Other data reduction functions may give slightly different results.
5. The concentration of the samples can be read directly from this standard curve. Samples with concentrations higher than that of the highest standard can be further diluted or must be reported as > 100 ng/mL. For the calculation of the concentrations this dilution factor must be considered.
6. For duplicate determinations, the mean of the two values must be taken. If the two values deviate substantially from one another DRG recommends retesting the samples.

Example of Typical Standard Curve

The following data is for demonstration only and cannot be used in place of data generations at the time of assay.

Standard

|

Optical Density (450 nm)

---|---
Standard 0 (0 ng/mL)|

0.02

Standard 1 (2 ng/mL)|

0.07

Standard 2 (5 ng/mL)|

0.16

Standard 3 (25 ng/mL)|

0.74

Standard 4 (50 ng /ml)|

1.41

Standard 5 (100 ng/mL)|

2.30

REFERENCE VALUES

It is strongly recommended that each laboratory determine its own normal and abnormal values.

In a study conducted with apparently healthy subjects, using the DRG Leptin Sandwich ELISA the following data were observed:

|

Adult females and males

---|---
BMI| 18.5 – 24.9| 25 – 30| > 30
n| 18| 12| 9
2.5th – 97.5th Percentile (ng/mL)| < 0.7 – 8.3| 1.5 – 19.3| 4.0 – 32.0
Mean (ng/mL)| 3.4| 7.4| 14.1
Median (ng/mL)| 2.3| 5.6| 7.1
Range (min. – max.) (ng/mL)| < 0.7 – 9.1| 1.3 – 21.2| 3.4 – 32.1

Children (1 – 10 years)

not determined

51

< 0.7 – 7.0

2.8

2.6

< 0.7 – 11.7

The results alone should not be the only reason for any therapeutic consequences. The results must be correlated to  other clinical observations and diagnostic tests.

QUALITY CONTROL

Good laboratory practice requires that controls be run with each calibration curve. A statistically significant number of controls should be assayed to establish mean values and acceptable ranges to assure proper performance.
It is recommended to use control samples according to state and federal regulations. The use of control samples is advised to assure the day-to-day validity of results. Use controls at both normal and pathological levels.
The controls and the corresponding results of the QC-Laboratory are stated in the Certificate of Analysis (CoA) added to the kit. The values and ranges stated on the Certificate of Analysis always refer to the current kit lot and must be used for direct comparison of the results.
If available, it is also recommended to make use of national or international Quality Assessment programs in order to ensure the accuracy of the results.
Employ appropriate statistical methods for analyzing control values and trends. If the results of the assay do not fit to the established acceptable ranges of control materials patient results should be considered invalid.
In this case, please check the following technical areas: Pipetting and timing devices; photometer, expiration dates of reagents, storage and incubation conditions, aspiration and washing methods.
After checking the above-mentioned items without finding any error contact your distributor or DRG directly.

PERFORMANCE CHARACTERISTICS

Measuring Range

The range of the assay is between 0.7 ng/mL – 100 ng/mL.

Specificity of Antibodies (Cross-Reactivity)

The following substances were tested for cross reactivity of the assay:

N.D.: Not detectable

Sensitivity

The analytical sensitivity [Mean OD(Standard 0) + 2 × SD, n = 20] was calculated to be 0.7 ng/mL.

Reproducibility

Intra-Assay

The within assay variability is shown below:

Sample

|

n

|

Mean (ng/mL)

|

CV (%)

---|---|---|---

1

|

20

|

1.1

|

9.6

2

|

20

|

3.2

|

7.8

3

|

20

|

27.4

|

8.6

Inter Assay

The between assay variability is shown below:

Sample

|

n

|

Mean (ng/mL)

|

CV (%)

---|---|---|---

1

|

6

|

1.4

|

6.9

2

|

6

|

3.7

|

3.7

3

|

6

|

9.7

|

9.1

Inter-Lot

The inter-assay (between-lots) variability is shown below:

Sample

|

n

|

Mean (ng/mL)

|

CV (%)

---|---|---|---

1

|

4

|

6.7

|

11.0

2

|

4

|

20.3

|

11.6

3

|

4

|

1.3

|

7.6

4

|

4

|

14.4

|

8.7

Recovery

Samples have been spiked by adding Leptin solutions with known concentrations.
The % recovery has been calculated by multiplication of the ratio of the measurements and the expected values with 100 (expected value = (endogenous Leptin + added Leptin) / 2; because of a 1:2 dilution of serum with spike material).

|

Sample 1

|

Sample 2

|

Sample 3

---|---|---|---

Concentration (ng/mL)

|

4.6

|

21.4

|

9.6

Average Recovery

|

88.8

|

97.0

|

94.8

Range of Recovery (%)  from to ****

|

86.8

|

90.6

|

84.0

93.1

|

102.1

|

106.0

Linearity

|

Sample 1

|

Sample 2

|

Sample 3

---|---|---|---

Concentration (ng/mL)

|

4.6

|

21.4

|

9.6

Average Recovery

|

93.2

|

92.7

|

104.7

Range of Recovery (%) from  to

|

85.1

|

86.2

|

88.0

107.5

|

103.1

|

114.3

LIMITATIONS OF USE

Reliable and reproducible results will be obtained when the assay procedure is performed with a complete understanding of the instructions for use and with adherence to good laboratory practice. Any improper handling of samples or modification of this test might influence the results.

Interfering Substances

Hemoglobin (up to 4 mg/mL), bilirubin (up to 0.5 mg/mL) and triglyceride (up to 30 mg/mL) have no influence on the assay results.
A biotin concentration of up to 1200 ng/mL in a sample has no influence on the assay results.

Drug Interferences

Until today no substances (drugs) are known to us, which have an influence on the measurement of leptin in a sample.

High Dose Hook Effect

“High Dose Hook Effect” is not detected up to 5000 ng/mL of leptin.

LEGAL ASPECTS

Reliability of Results

The test must be performed exactly as per the manufacturer’s instructions for use. Moreover, the user must strictly adhere to the rules of GLP (Good Laboratory Practice) or other applicable national standards and/or laws. This is especially relevant for the use of control reagents. It is important to always include, within the test procedure, a sufficient number of controls for validating the accuracy and precision of the test.
The test results are valid only if all controls are within the specified ranges and if all other test parameters are also within the given assay specifications. In case of any doubt or concern please contact DRG.

Therapeutic Consequences

Therapeutic consequences should never be based on laboratory results alone even if all test results are in agreement with the items as stated under point 11.1. Any laboratory result is only a part of the total clinical picture of a patient.
Only in cases where the laboratory results are in acceptable agreement with the overall clinical picture of the patient should therapeutic consequences be derived.
The test result itself should never be the sole determinant for deriving any therapeutic consequences.

Liability

Any modification of the test kit and/or exchange or mixture of any components of different lots from one test kit to another could negatively affect the intended results and validity of the overall test. Such modification and/or exchanges invalidate any claim for replacement.
Claims submitted due to customer misinterpretation of laboratory results subject to point 11.2 are also invalid.
Regardless, in the event of any claim, the manufacturer’s liability is not to exceed the value of the test kit. Any damage caused to the test kit during transportation is not subject to the liability of the manufacturer.

Reporting of Serious Incident

Any serious incident that has occurred in relation to the device shall be reported to the manufacturer and the competent authority of the Member State in which the user and/or the patient is established.

REFERENCES / LITERATURE

1. Londraville RL, et al. Comparative endocrinology of leptin: assessing function in a phylogenetic context. Gen Comp Endocrinol (2014) 203:146–57.
2. Bjorbaek C, Kahn BB. Leptin signaling in the central nervous system and the periphery. Recent Prog Horm Res (2004) 59:305–32.
3. Arora S. Leptin and its metabolic interactions – an update. Diabetes Obes Metab (2008) 10(11): 973–93.
4. Elias CF et al. Leptin differentially regulates NPY and POMC neurons projecting to the lateral hypothalamic area. Neuron (1999) 23(4):775–86.
5. Elmquist JK. Hypothalamic pathways underlying the endocrine, autonomic, and behavioral effects of leptin. Physiol Behav (2001) 74(4):703–8.
6. Brennan AM, Mantzoros CS. Drug Insight: the role of leptin in human physiology and pathophysiology–emerging clinical applications”. Nat Clin Pract Endocrinol Metab. (2006) 2 (6): 318–27.
7. Considine RV et al. “Serum immunoreactive-leptin concentrations in normal-weight and obese humans”. N Engulf Med. (1996). 334 (5): 292–5.
8. Pan H, Guo J, Su Z “Advances in understanding the interrelations between leptin resistance and obesity”. Physiology & Behavior. (2014) 130: 157–169.
9. Ahima RS, Qi Y, Singhal NS, Jackson MB, Scherer PE. Brain adipocytokine action and metabolic regulation. Diabetes. (2006) 55(Suppl 2):145–54.
10. Ahima RS, Flier JS. Adipose tissue as an endocrine organ. Trends Endocrinol Metab. (2000) 11(8): 327–32.
11. Weigle DS et al. Recombinant ob protein reduces feeding and body weight in the ob/ob mouse. J Clin Invest. (1995) 96(4):2065
12. Farooqi IS et al. Beneficial effects of leptin on obesity, T cell hyporesponsiveness, and neuroendocrine/metabolic dysfunction of human congenital leptin deficiency.  J Clin Invest. (2002) 110(8):1093–103.
13. Ma Z, et al. Radioimmunoassay of leptin in human plasma Clin Chem. (1996); 42 (6 Pt 1): 942-6
14. Wallace AM. Measurement of Leptin and Leptin binding in the human circulation Ann of Clin Biochem. (2000), 37: 244-52
15. Meier et al. Endocrine regulation of energy metabolism: a review of path biochemical and clinical chemical aspects of leptin, ghrelin, adiponectin, and resistin Clin Chem. (2004); 50 (9): 1511-25

| European Conformity
---|---
| Consult instructions for use •
IVD| In vitro diagnostic medical device”

REF

| Catalogue number •
LOT| Batch code•

| Contains sufficient for tests ..
| Temperature limit •
| Use-by date •
| Manufacturer”
| Distributed by
| Biological risks •
| Caution”‘
|
| F o r r esea r c h u se only

Content

| Content

Volume / No .

| Volume / No.
|

Microtiter wells

| Microterwells

Antiserum

| Antiserum

Enzyme Conjugate

| Enzyme Conjugate

Enzyme Complex

| Enzyme Complex

Substrate Solution

| Substrate Solution

Stop Solution

| Stop Solution

Zero Standard

| Zero Standard

Standard

| Standard

Control

| Control

Assay Buffer

| Assay Buffer

Wash Solution

| Wash Solution

1NNaOH

| 1N NaOH

1 N HGI

| 1 N HCI

Sample Diluent

| Sample Diluent

Conjugate Diluent

| Conjugate Diluent

Customer Support

EIA-2395
96
DRG Instruments GmbH, Germany Frauen berg strabe 18, 35039 Marburg
Phone: +49 (0)6421-1700 0, Fax: +49 (0)6421-1700 50
Website: www.drg-diagnostics.de 
E-mail: drg@drg-diagnostics.de
DRG International, Inc., USA
841 Mountain Ave., Springfield, NJ 07081
Phone: 973-564-7555, Fax: 973-564-7556
Website: www.drg-international.com
E-mail: corp@drg-international.com

References

• Analyticon® Biotechnologies GmbH

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