Sample Typle

Plasma,Serum,Cell culture supernatant, other body fluid

Contents

Standard Curve

Sensitivity

8µM-400µM

Reaction Time

15 minutes

Applications

For biological research: Glucose measurement in biological samples For drug/pharm research: Drug influence on Glucose metabolism

Storage

-80°C

Materials needed but not supplied

A plate reader capable of measuring absorbance at 490nm

Adjustable pipettes and a repeat pipettor

Distilled water(milliQ or HPLC-grade)

0.5M Acetic Acid

Clear flat bottom 96-well plates if not included in the kit purchased

1. Glucose Standard(2x)

Note: Please don’t do dilution in the provided Glucose standard vial itself.

The vial contains 500µl of 800µM Glucose Standard. The standard must be equilibrated to room temperature before use. Dilute 500µl of 800µM Glucose Standard with 500µl of dH2O to prepare a 400μM Glucose Standard. 1ml of diluted standard is enough for making 3 standard curves if assayed in duplicate. Store at -80?C.

2. Glucose Assay Solution

The solution contains enzymes that are light sensitive. The solution must be thawed on ice before use.Best to aliquot the amount needed and use it all to prevent thawing/freezing cycles. Freeze and store aliquots at -80 oC

1.Sample Preparation

Serum/Plasma/other body fluid/cell culture supernatant Serum, Plasma, other body fluid, or cell culture supernatant can be measured directly by a series of dilutions of the sample (1/2; 1/4; 1/8; .…..) to ensure the readings are within the standard curve. Your samples can be diluted with dH2O. Solid Samples Solid samples, such as tissues, can be first homogenized and extracted with ethanol (80%) with a tissues/Ethanol ratio of 1:8 (1 hr at 4°C) followed by centrifugation at 10,000xg. The clear supernatants then can be measured directly by a series of dilutions of the sample (1/2; 1/4; 1/8; .…..) to ensure the readings are within the standard curve. Your samples can be diluted with dH2O. Adding Samples Add 50μl of samples to each well. We recommend that samples be assayed in duplicate.

Note: If prepared samples are not assayed the same day, store the samples at -80?C. The samples will be stable for one month while stored at -80?C. For frozen samples, dilutions of samples must be done right before assaying.

We recommend that Glucose Standards be assayed in duplicate. A standard curve has to be run in each assay. Add 50µl, 40µl, 30µl, 20µl, 10µl, 5µl, 1 µl, and 0µl of diluted Glucose Standard to each well. Then adjust volume to 50μl/well with dH2O.

3.Perform the assay a. Add 50µl of Glucose Assay Solution to each well containing the Glucose Standards and test samples. b. Incubate for 15mins at 37°C incubator. Note: Please do not use CO2 incubator. c. Stop the reaction by adding 50μl of 0.5 M acetic acid per well followed by brief gentle agitation. Note: Eliminate any air bubbles present in the wells using a needle prior to measurement. d. Measure the absorbance at 490nm using a microplate reader.

4. Calculation

a. Average the OD490 nm values of replicate wells of each Glucose standard, test samples, and blank. In order to get the corrected absorbance, subtract the average OD490 nm value of the blank from the averaged OD490 nm values from all standards and samples. b. Make a standard curve by plotting OD490 nm values from each glucose standards as a function of Glucose concentration. This can be done with excel spreadsheet. Calculate the concentration of Glucose in samples using the equation obtained from the linear regression of the standard curve.

Material Safety Data Sheet

Date:Oct 26,2015

1. PRODUCT AND COMPANY IDENTIFICATION

1.1 Product identifier

1.2 Relevant identified uses of the substance or mixture and uses advised against

1.3 Details of the supplier of the safety data sheet

1.4 Emergency telephone number

2. HAZARDS IDENTIFICATION

2.1 Classification of the substance or mixture

2.2 GHS Label elements, including precautionary statements

2.3 Other hazards

None

3. COMPOSITION/INFORMATION ON INGREDIENTS

3.1 Substances

4. FIRST AID MEASURES

4.1 Description of first aid measures

Eye contact

Skin Contact

Inhalation

Ingestion

4.2 Most important symptoms and effects, both acute and delayed

4.3 Indication of any immediate medical attention and special treatment needed.

5. FIRE FIGHTING MEASURES

5.1 Extinguishing media

Suitable extinguishing media

5.2 Special hazards arising from the substance or mixture

5.3 Advice for firefighters

6. ACCIDENTAL RELEASE MEASURES

6.1 Personal precautions,protective equipment and emergency procedures

6.2 Environmental precautions

6.3 Methods and material for containment and cleaning up

7. HANDLING AND STORAGE

7.1 Precautions for safe handling

7.2 Conditions for safe storage,including any incompatibilities

7.3 Specific end use(s)

8. EXPOSURE CONTROLS/PERSONAL INFORMATION

8.1 Control Parameters

8.2 Exposure controls

Engineering controls

Personal protective equipment

9. PHYSICAL AND CHEMICAL PROPERTIES

9.1 Information on basic physical and chemical properties

9.2 Other safety information

10. STABILITY AND REACTIVITY

10.1Reactivity

10.2Chemical stability

10.3Possibility of hazardous reactions

10.4Conditions to avoid

10.5Incompatible materials

10.6Hazardous decomposition products

11. TOXICOLOGICAL INFORMATION

11.1Information on toxicological effects

12. ECOLOGICAL INFORMATION

12.1Toxicity

12.2Persistence and degradability

12.3.Bioaccumulative potential

12.4Mobility in soil

12.5Results of PBT and vPvB assessment

12.6Other adverse effects

13. DISPOSAL INFORMATION

13.1Waste treatment methods

14. TRANSPORT INFORMATION

15. REGULATORY INFORMATION

16. OTHER INFORMATION

It is Glucose Oxidase.

Unfortunately it is proprietary.

It is 8μM-400μM.

Yes it is necessary since you calculate your sample concentration based on the standard curve. Please do not use the representative standard curve in the protocol to calculate concentrations of your samples.

End-users would have to perform an experiment to determine optimal volume of samples whose readings fall within the standard curve range. Please refer to publications with which this product was used to see how much samples have been used in similar experiments.

Your samples may be diluted with PBS or dH2O.

No, the kit does not work with a fluorescence reader since the kit employs colorimetric assays.

Although it is better to use fresh samples for assays, you can use frozen samples. If you are not using frozen samples in once, please make aliquots of your samples before you put them in a freezer to prevent from degrading samples from repeated freeze-thaw cycles.

You can still measure readings without adding acetic acid.

It is ok as long as you set up the absorbance between 470 and 490nm.

Many factors influence the values such as room temperature, incubation time, handling, etc.

Yes they are available for first time customers; however, you will be responsible for shipping and handling fee. Please contact