Glucose Assay Kit I

Glucose is the most important carbohydrate in biology. Transported via the blood stream, it is the primary source of energy for the body’s cells. Glucose levels are highly regulated in the human body. Failure to maintain blood glucose in the normal range leads to conditions of persistently high or low blood sugar. Diabetes mellitus is the most prominent disease related to improper blood sugar regulation. The determination of glucose levels in blood is critical in the control of diabetes. Eton’s Glucose Assay Kit provides a simple, reproducible, and sensitive tool for assay glucose in plasma, serum, and urine. The glucose assay uses the glucose oxidase-peroxide reaction for the determination of glucose concentrations. The color intensity of the reaction product at 490nm is directly proportional to the glucose concentration in the sample. The kit is stable until the expiration date under proper storage and handling conditions. The product is for research only.

Sku# Product Name Product Size Price QTY

Sku#

120003100P

Product Name

Glucose Assay Kit I (5ml)-100 Assays w/Reader Plate(1x)

Product Size

(5ml) - 100 assays w/Reader Plate (1x)

Price

$132.00 USD

QTY

Sku#

120003100A

Product Name

Glucose Assay Kit I (5 ml)-100 Assays- Assay Solution ONLY

Product Size

(5 ml)-100 Assays- Assay Solution ONLY

Price

$107.80 USD

QTY

Sku#

1200031002

Product Name

Glucose Assay Kit I (5 ml) -100 Assays

Product Size

(5 ml) -100 Assays

Price

$118.80 USD

QTY

Sku#

120003400P

Product Name

Glucose Assay Kit I (20ml)-400 Assays w/Reader Plate(4x)

Product Size

(20ml) - 400 assays w/Reader Plate (4x)

Price

$358.60 USD

QTY

Sku#

120003400A

Product Name

Glucose Assay Kit I (20ml)-400 Assays- Assay Solution ONLY

Product Size

(20 ml)-400 Assays- Assay Solution ONLY

Price

$293.70 USD

QTY

Sku#

1200034002

Product Name

Glucose Assay Kit I (20 ml) -400 Assays

Product Size

(20 ml) -400 Assays

Price

$304.70 USD

QTY

Sku#

120003200P

Product Name

Glucose Assay Kit I (10ml)-200 Assays w/Reader Plate(2x)

Product Size

(10ml) - 200 assays w/Reader Plate (2x)

Price

$198.00 USD

QTY

Sku#

120003200A

Product Name

Glucose Assay Kit I (10 ml)-200 Assays- Assay Solution ONLY

Product Size

(10 ml)-200 Assays- Assay Solution ONLY

Price

$160.60 USD

QTY

Sku#

1200032002

Product Name

Glucose Assay Kit I (10 ml) -200 Assays

Product Size

(10 ml) -200 Assays

Price

$171.60 USD

QTY

Add to Cart

Sample Typle

Plasma,Serum,Cell culture supernatant, other body fluid

Contents



Method

Colorimetric method at 490nm

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

Notes

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

Shipping Icepacks


Reagent Preparation

Note: All reagents are frozen. We recommend you spin the standard vial before opening.

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

Protocol

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.

2.Standard Curve Prepation

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.

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 I

Catalog No: 1200031002, 120003100A, 120003100P, 1200032002, 120003200A, 12003200P, 1200034002, 120003400A, 120003400P

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 Solution

Components

CAS#

Potassium phosphate monobasic

7778-77-0

Potassium phosphate dibasic

7758-11-4

Glucose Oxidase

9001-37-0

Glucose Assay Standard

Components

CAS#

Sodium D-Glucose

50-99-7

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

Ingestion

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

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

Appearance

Clear, colorless liquid

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

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.



Zakikhani M, Bazile M, Hashemi S, Javeshghani S, Avizonis D, et al. (2012) Alterations in Cellular Energy Metabolism Associated with the Antiproliferative Effects of the ATM Inhibitor KU-55933 and with Metformin. PLoS ONE 7(11): e49513. doi:10.1371/journal.pone.0049513
Shuang Liang,* Paola Galluzzo,* Anna Sobol, Sylvia Skucha, Brittany Rambo, and Maurizio Bocchetta.Multimodality Approaches to Treat Hypoxic Non–Small Cell Lung Cancer (NSCLC) Microenvironment.Genes Cancer. 2012 February; 3(2): 141–151.
ZHANG, YANBIN et al. Glucose Transporter 3 Performs a Critical Role in mTOR-Mediated Oncogenic Glycolysis and Tumorigenesis.Oncology Letters 9.6 (2015): 2809–2814. PMC. Web. 6 Oct. 2015.
Eyal Amiel,Bart Everts,Tori C.Freitas,Irah L.King,Jonathan D.Curtis, Erika L.Pearce and Edward J.Pearce.Inhibition of Mechanistic Target of Rapamycin Promotes Dendritic Cell Activation and Enhances Therapeutic Autologous Vaccination in Mice.J Immunol 2012;189:2151-2158
Zhiguo Li,Jie Li,Penpeng Bi,Ying Lu,Grant Burcham, Bennet D.Elzey,Timothy Ratliff,Stephen F.Konieczny,Nihal Ahmad,Shihuan Kuang,and Xiaoqui Liu.Plk1 Phosphorylation of PTEN causes a tumor-Promoting Metabolic State.Mol. Cell. Biol. October 2014 vol. 34 no. 19 3642-3661

Qingsong Cai,Tong Lin, Sushama Kamarajugadda, and Jianrong Lu. Regulation of Glycolysis and the Warburg Effect by Estrogen-related Receptors.Oncogene. 2013 April 18; 32(16): 2079–2086.




What type of medium should I use for making cultured cells for this assay?
Please use phenol red free medium. Please do not use phenol red medium since phenol red would affect absorbance readings.

What enzyme is used in this assay?

It is Glucose Oxidase.

What are the compositions of this assay?

Unfortunately it is proprietary.

What is the sensitivity of this assay?

It is 8μM-400μM.

Do I need to make a standard curve every time?

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.

How much sample do I need for this assay?

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.

How can I dilute my samples?

Your samples may be diluted with PBS or dH2O.

Can I use fluorescence spectroscopy to measure readings for this assay?

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

Can I use frozen samples?

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.

My sample formed precipitation after adding acetic acid. How can I prevent this?

You can still measure readings without adding acetic acid.

The protocol recommends measuring the absorbance at 490nm. Is 485nm close enough?

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

Why are my standard curve values different than the one shown on the datasheet?

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

Are trial kits available?

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

for more details.


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