Sample Typle	Plasma,Serum,Cell culture supernatant, other body fluid
Components
Method	Colorimetric method at 570nm
Standard Curve
Sensitivity	4µM-200µM
Reaction Time	30 minutes
Applications	For biological research: Glucose measurement in biological samples For drug/pharm research: Drug influence on Glucose metabolism
Storage	-80°C
Notes	Material needed but not supplied A plate reader capable of measuring absorbance at 570nm Adjustable pipettes and a repeat pipettor Distilled water(milliQ or HPLC-grade) Clear Flat bottom 96-well plates if not included in the kit purchased
Shipping	Icepacks

Reagent Preparation Note: All reagents are frozen. We recommend you spin small vials before opening. 1. Glucose Assay Buffer(10x) Note: Please don’t do dilution in the provided Glucose Assay Buffer bottle itself. The bottle contains 10ml of 10x Glucose Assay Buffer. The assay buffer must be equilibrated to room temperature. Please make appropriate amount of 1xGlucose Assay Buffer for the assay, for example, by diluting 1ml of 10xGlucose Assay Buffer with 9 ml of distilled water. Prepare additional diluted assay buffer if needed. Store at -80?C. 2. Glucose Standard (10x) Note: Please don’t do dilution in the provided Glucose Standard vial itself. The vial contains 100µl of 2mM Glucose Standard. The standard must be equilibrated to room temperature before use. Dilute 100µl of 2mM Glucose Standard with 900µl of diluted Glucose Assay Buffer to prepare a 200μM Glucose Standard. 1ml of diluted Glucose Standard is enough for making 3 standard curves if assayed in duplicate. Store at -80?C. 3. Assay Probe The vial contains 45μl of Assay Probe. The assay probe must be equilibrated to room temperature before use. It is sufficient for 100 wells.Protect from light. Store at -80?C. 4. Glucose Enzyme Mix The vial contains 70μl of Glucose Enzyme Mix. The enzyme mix must be thawed on ice before use. It is sufficient for 100 wells. Store at -80?C.

Protocol 1.Sample Preparation Blood (serum/plasma), urine, other body fluids 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 range.Your samples can be diluted with dH2O or diluted Glucose Assay Buffer. 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.

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

3.Reaction Solution Preparation 50μl Reaction Mix is required for each well and 1ml of Reaction Solution is prepared with the followings: 980µlGlucose Assay Buffer(1x) 12µlGlucose Assay Enzyme Mix 8µlAssay Probe 1ml of Reaction Solution is enough for ~20 wells. Prepare appropriate amount of Reaction Solution for the number of assays to be performed. Note:Please do not keep any leftover Reaction Solution as it has to be made fresh right beforeyou run assays. If old Reaction Solution is used, assays will not work.

4.Perform the assay a. Mix the reaction solution well and add 50μl of the reaction solution to each well containing the Glucose standards and test samples. b. Incubate the reaction for 30 minutes in a 37?C incubator. c. Measure absorbance at 570 nm in a microplate reader.

5. Calculation a. Average the OD570 nm values of replicate wells of each Glucose Standard, test samples, and blank. Then, correct background by subtracting the value derived from the blank from all sample readings. b. Make a standard curve by plotting OD570 nm values from each Glucose Standards as a function of Glucose concentration. This can be done with excel spreadsheet. c. Calculate the value of Glucose in samples using the equation obtained from the linear regression of the standard curve. Glucose (μM)= [ (Corrected absorbance)-(y-intercept)]/Slope

Material Safety Data Sheet

Date:Sept 21,2017

1. PRODUCT AND COMPANY IDENTIFICATION

1.1 Product identifier

Product name: Glucose Assay Kit II

Catalog No: 1200071002, 120007100P, 1200072002, 120007200P

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

For research use only.

1.3 Details of the supplier of the safety data sheet

Company:Eton Bioscience Inc

TollFree:1-800-758-1630

Tel:1-800-758-1630

Fax:1-800-507-2912

1.4 Emergency telephone number

1-800-758-1630

2. HAZARDS IDENTIFICATION

2.1 Classification of the substance or mixture

Not a hazardous substance or mixture

2.2 GHS Label elements, including precautionary statements

Not a hazardous substance or mixture

2.3 Other hazards

None

3. COMPOSITION/INFORMATION ON INGREDIENTS

3.1 Substances

	Glucose Assay Buffer				Glucose Assay Enzyme Mix
	Components		CAS#		Components	CAS#
	Potassium Phosphate Monobasic		7778-77-0		Potassium Phosphate Monobasic	7778-77-0
	Potassium Phosphate Dibasic		7758-11-4		Potassium Phosphate Dibasic	7758-11-4
	Water		7732-18-5		Glucose Oxidase	9001-37-0
	Glucose Standard				Assay Probe
	Components		CAS#		Components	CAS#
	Sodium D-Glucose		50-99-7		DMSO	67-68-5
	Potassium Phosphate Monobasic		7778-77-0
	Potassium Phosphate Dibasic		7758-11-4
	Water		7732-18-5

4. FIRST AID MEASURES

4.1 Description of first aid measures

Eye contact

Check for and remove contact lenses and flush with copious amounts of water; assure adequate flushing by separating the eyelids with fingers; call a physician

Skin Contact

Flush with copious amounts of water; remove contaminated clothing and shoes; call a physician

Inhalation

Remove to fresh air. If not breathing give artificial respiration ;call a physician

Ingestion

If swallowed, never give anything by mouth to an unconscious person. Rinse mouth with water ;call a physician

4.2 Most important symptoms and effects, both acute and delayed

No information available

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

No information available

5. FIRE FIGHTING MEASURES

5.1 Extinguishing media

Suitable extinguishing media

Water spray, alcohol-resistant foam, dry chemical or carbon dioxide

5.2 Special hazards arising from the substance or mixture

No information available

5.3 Advice for firefighters

Wear self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes.

6. ACCIDENTAL RELEASE MEASURES

6.1 Personal precautions, protective equipment and emergency procedures

Use full personal protective equipment. Evacuate personnel to safe areas. Avoid breathing vapors,mist or gas.

6.2 Environmental precautions

Prevent further leakage or spillage. Prevent product from entering drain.

6.3 Methods and material for containment and cleaning up

Keep in suitable, closed containers for disposal

7. HANDLING AND STORAGE

7.1 Precautions for safe handling

Avoid skin/eye contact. Use protective equipment as needed. Wash contaminated clothing before reuse.

7.2 Conditions for safe storage,including any incompatibilities

Keep container tightly closed. Store at -80?C.

7.3 Specific end use(s)

No data available.

8. EXPOSURE CONTROLS/PERSONAL INFORMATION

8.1 Control Parameters

This product contains no hazardous materials with occupational exposure limits.

8.2 Exposure controls

Engineering controls

Provide shower and eye wash station

Personal protective equipment

Eye Protection	Wear safety goggles
Skin Protection	Wear protective clothing
Hand Protection	Wear protective gloves
Respiratory protection	Wear NIOSH/MSHA approved respirators

9. PHYSICAL AND CHEMICAL PROPERTIES

9.1 Information on basic physical and chemical properties

Glucose Assay Buffer

Appearance	Liquid, colorless
pH	7.0
Melting point	No data available
Freezing point	No data available
Boiling point	No data available
Density	No data available
Water solubility	Soluble

Glucose Standard

Appearance	Liquid, colorless
pH	7.0
Melting point	No data available
Freezing point	No data available
Boiling point	No data available
Density	No data available
Water solubility	Soluble

Glucose Assay Enzyme Mix

Appearance	Liquid, yellow
pH	No data available
Melting point	No data available
Freezing point	No data available
Boiling point	No data available
Density	No data available
Water solubility	Soluble

Assay Probe

Appearance	Liquid, colorless
pH	No data available
Melting point	No data available
Freezing point	No data available
Boiling point	No data available
Density	No data available
Water solubility	Soluble

9.2 Other safety information

No data available.

10. STABILITY AND REACTIVITY

10.1Reactivity

No data available

10.2Chemical stability

Stable under recommended storage conditions.

10.3Possibility of hazardous reactions

No data available.

10.4Conditions to avoid

No data available

10.5Incompatible materials

Strong oxidizing agents

10.6Hazardous decomposition products

Hazardous decomposition products formed under fire conditions: carbon oxides

11. TOXICOLOGICAL INFORMATION

11.1Information on toxicological effects

Toxicological effects on this product have not been thoroughly studied.

12. ECOLOGICAL INFORMATION

12.1Toxicity

Avoid release into environment

12.2Persistence and degradability

No data available

12.3.Bioaccumulative potential

No data available

12.4Mobility in soil

No data available

12.5Results of PBT and vPvB assessment

No data available

12.6Other adverse effects

No data available

13. DISPOSAL INFORMATION

13.1Waste treatment methods

Dispose in accordance with local, state, and federal regulations.

14. TRANSPORT INFORMATION

DOT:

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport
IATA:

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport

ADR/RID

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport

15. REGULATORY INFORMATION

EU Risk and Safety phrases:

R36/37/38: irritating to eyes/respiratory system/skin

16. OTHER INFORMATION

The information above is believed to be accurate and represents the best information currently available to us. However, we make no warranty of merchantability or any other warranty, express or implied, with respect to such information. Eton Bioscience Inc. shall not be held liable for any damages or other consequences resulting from handling or from contact with the above product.