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OUR SUPPLIERS
KOMABIOTECH
301,
Gayang Technotown, #1487
Gayang 3 dong,
Gangseo-gu
Seoul 157-793, KOREA
Spherotech, Inc.
27845 Irma Lee Circle, Unit 101
Lake Forest, IL 60045
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Exalpha
Biologicals,
Inc.
2 Shaker Road,
Unit B101
Shirley, MA
01464
SCETI K.K
BIOSCIENCE Export DF Kasumigaseki Place,3-6-7 Kasumigaseki Chiyoda-ku, Tokyo 100-0013 JAPAN
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EY
Laboratories, Inc. Headquarters
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San Mateo, CA. 94401 USA
EXBIO Praha, a.s.
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Czech Republic
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0032 (0)16 41 44 07 |
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QuantiChromTM Glucose Assay
Kit
Quantitative Colorimetric
Glucose Determination at 630nm
DESCRIPTION
Glucose (C6H12O6) is a
ubiquitous fuel molecule in biology. It is
oxidized through a series of enzyme-catalyzed
reactions to form carbon dioxide and water,
yielding the universal energy molecule ATP. Due
to its importance in metabolism, glucose level
is a key diagnostic parameter for many metabolic
disorders. Increased glucose levels have been
associated with diabetes mellitus, hyperactivity
of thyroid, pituitary and adrenal glands.
Decreased levels are found in insulin secreting
tumors, myxedema, hypopituitarism and
hypoadrenalism. Simple, direct and
automation-ready procedures for measuring
glucose concentrations find wide applications in
research and drug discovery. BioAssay Systems'
glucose assay kit is designed to measure glucose
directly in serum or plasma without any
pretreatment. The improved otoluidine
method utilizes a specific color reaction with
glucose. The absorbance at 630nm is directly
proportional to glucose concentration in the
sample.
KEY FEATURES
Sensitive and accurate .
Use as little as 5 μL samples. Linear detection
range 0.7 mg/dL (39 μM) to 300 mg/dL (16.6 mM)
glucose in 96-well plate.
Simple and convenient. The
procedure involves addition of a single working
reagent and incubation for 8 min in a boiling
water bath.
Improved reagent stability.
The optimized formulation has greatly enhanced
the reagent and signal stability.
Low interference in biological
samples. No pretreatments are needed. Assays
can be directly performed on serum and plasma
samples.
APPLICATIONS:
Direct Assays:
glucose in biological samples (e.g. serum and
plasma).
Drug Discovery/Pharmacology:
effects of drugs on glucose metabolism.
Food and Beverages: glucose in food,
beverages etc.
Storage conditions. The
kit is shipped at room temperature. Store the
reagent at room temperature and standard at
-20°C, respectively. Shelf life of at least 6
months (see expiry dates on labels).
Precautions: reagents are
for research use only. Normal precautions for
laboratory reagents should be exercised while
using the reagents. Please refer to Material
Safety Data Sheet for detailed information.
PROCEDURES
IMPORTANT: THE REAGENT
CONTAINS ACETIC ACID. THIS ASSAY IS PREFERABLY
CARRIED OUT IN A CHEMICAL FUME HOOD.
Procedure using 96-well
plate:
1. Dilute standard in
distilled water as follows.
Set up 1.5-mL centrifuge
tubes. Transfer 5 μL diluted standards and
samples to appropriately labeled tubes. Transfer
500 μL Reagent to each tube. Close the tubes
tightly and mix. Store diluted standards at
-20°C for future use.
2. Place the tubes in a tube
holder and heat in a boiling water bath or heat
block for 8 min. Cool down in cold water bath
for 4 min.
3. Transfer 200 μL in
duplicate into a clear bottom 96-well plate.
Careful: avoid bubble formation. Read optical
density at 620-650nm (peak absorbance at 630nm).
Procedure using cuvette:
1. Dilute standards and
transfer 12 μL water blank, Standards and
samples to appropriately labeled tubes. Transfer
1200 μL Reagent to each tube. Close the tubes
tightly and mix.
2. Place the tubes in a tube
holder and heat in a boiling water bath for 8
min. Cool down in cold- water bath for 4 min.
3. Transfer 1000
μL
reaction mixture into cuvet. Read optical
density at 620-650nm (peak absorbance at 630nm)
against blank.
Note :
1. if the Sample OD is higher than the Standard
OD at 300mg/dL, dilute sample in water and
repeat the assay.
2. To determine low glucose
concentrations, use 50
μL sample and standards
(instead of 5 μL) per 500 μL
Reagent.
CALCULATION
Subtract blank OD (water, #5)
from the standard OD values and plot the OD
against standard concentrations. Determine the
slope using linear regression fitting. The
glucose concentration of Sample is calculated as
OD SAMPLE
and ODBLANK
are
optical density values of the sample and sample
“Blank” (water or buffer in which the sample was
diluted). Typical serum/plasma glucose values:
70 - 110 mg/dL.
Conversions :
1mg/dL glucose equals 55.5 μM, 0.001% or 10 ppm.
MATERIALS REQUIRED, BUT NOT
PROVIDED
Pipeting devices, centrifuge
tubes, boiling water bath, tube holder.
Procedure using 96-well
plate:
Clear bottom 96-well plates
(e.g. Corning Costar) and plate reader.
Procedure using cuvette:
Spectrophotometer and Cuvets
for measuring OD at 620-650nm.
EXAMPLES:
Rat plasma, rat serum, goat
serum and human plasma were assayed using the
96-well plate assay protocol. The glucose
concentrations were 128
± 2 (n = 4), 72.5 ± 0.8 (n = 4), 78.6 ±
0.6, 69.3
± 0.7 mg/dL (n = 4), respectively. Coefficient
of variance < 3%.
PUBLICATIONS
1. Yoon, S.S. and Mekalanos,
J.J. (2006) 2,3-Butanediol synthesis and the
emergence of the
Vibrio
cholerae El Tor biotype. Infection and
Immunity 74 (12): 6547–6556.
2. Schmidt, C. et al (2007).
Regulation of renal glucose transporters during
severe inflammation. Am J Physiol Renal Physiol
292: F804- F811.
3. Jatana, M. (2006)
Inhibition of NF- κB
activation by 5- lipoxygenase inhibitors
protects brain against injury in a rat model of
focal cerebral ischemia. J. Neuroinflammation
3:12. |
|
QuantiChromTM Glucose Assay
Kit
Quantitative Colorimetric
Glucose Determination at 630nm
DESCRIPTION
Glucose (C6H12O6) is a
ubiquitous fuel molecule in biology. It is
oxidized through a series of enzyme-catalyzed
reactions to form carbon dioxide and water,
yielding the universal energy molecule ATP. Due
to its importance in metabolism, glucose level
is a key diagnostic parameter for many metabolic
disorders. Increased glucose levels have been
associated with diabetes mellitus, hyperactivity
of thyroid, pituitary and adrenal glands.
Decreased levels are found in insulin secreting
tumors, myxedema, hypopituitarism and
hypoadrenalism. Simple, direct and
automation-ready procedures for measuring
glucose concentrations find wide applications in
research and drug discovery. BioAssay Systems'
glucose assay kit is designed to measure glucose
directly in serum or plasma without any
pretreatment. The improved otoluidine
method utilizes a specific color reaction with
glucose. The absorbance at 630nm is directly
proportional to glucose concentration in the
sample.
KEY FEATURES
Sensitive and accurate .
Use as little as 5 μL samples. Linear detection
range 0.7 mg/dL (39 μM) to 300 mg/dL (16.6 mM)
glucose in 96-well plate.
Simple and convenient. The
procedure involves addition of a single working
reagent and incubation for 8 min in a boiling
water bath.
Improved reagent stability.
The optimized formulation has greatly enhanced
the reagent and signal stability.
Low interference in biological
samples. No pretreatments are needed. Assays
can be directly performed on serum and plasma
samples.
APPLICATIONS:
Direct Assays:
glucose in biological samples (e.g. serum and
plasma).
Drug Discovery/Pharmacology:
effects of drugs on glucose metabolism.
Food and Beverages: glucose in food,
beverages etc.
Storage conditions. The
kit is shipped at room temperature. Store the
reagent at room temperature and standard at
-20°C, respectively. Shelf life of at least 6
months (see expiry dates on labels).
Precautions: reagents are
for research use only. Normal precautions for
laboratory reagents should be exercised while
using the reagents. Please refer to Material
Safety Data Sheet for detailed information.
PROCEDURES
IMPORTANT: THE REAGENT
CONTAINS ACETIC ACID. THIS ASSAY IS PREFERABLY
CARRIED OUT IN A CHEMICAL FUME HOOD.
Procedure using 96-well
plate:
1. Dilute standard in
distilled water as follows.
Set up 1.5-mL centrifuge
tubes. Transfer 5 μL diluted standards and
samples to appropriately labeled tubes. Transfer
500 μL Reagent to each tube. Close the tubes
tightly and mix. Store diluted standards at
-20°C for future use.
2. Place the tubes in a tube
holder and heat in a boiling water bath or heat
block for 8 min. Cool down in cold water bath
for 4 min.
3. Transfer 200 μL in
duplicate into a clear bottom 96-well plate.
Careful: avoid bubble formation. Read optical
density at 620-650nm (peak absorbance at 630nm).
Procedure using cuvette:
1. Dilute standards and
transfer 12 μL water blank, Standards and
samples to appropriately labeled tubes. Transfer
1200 μL Reagent to each tube. Close the tubes
tightly and mix.
2. Place the tubes in a tube
holder and heat in a boiling water bath for 8
min. Cool down in cold- water bath for 4 min.
3. Transfer 1000
μL
reaction mixture into cuvet. Read optical
density at 620-650nm (peak absorbance at 630nm)
against blank.
Note :
1. if the Sample OD is higher than the Standard
OD at 300mg/dL, dilute sample in water and
repeat the assay.
2. To determine low glucose
concentrations, use 50
μL sample and standards
(instead of 5 μL) per 500 μL
Reagent.
CALCULATION
Subtract blank OD (water, #5)
from the standard OD values and plot the OD
against standard concentrations. Determine the
slope using linear regression fitting. The
glucose concentration of Sample is calculated as
OD SAMPLE
and ODBLANK
are
optical density values of the sample and sample
“Blank” (water or buffer in which the sample was
diluted). Typical serum/plasma glucose values:
70 - 110 mg/dL.
Conversions :
1mg/dL glucose equals 55.5 μM, 0.001% or 10 ppm.
MATERIALS REQUIRED, BUT NOT
PROVIDED
Pipeting devices, centrifuge
tubes, boiling water bath, tube holder.
Procedure using 96-well
plate:
Clear bottom 96-well plates
(e.g. Corning Costar) and plate reader.
Procedure using cuvette:
Spectrophotometer and Cuvets
for measuring OD at 620-650nm.
EXAMPLES:
Rat plasma, rat serum, goat
serum and human plasma were assayed using the
96-well plate assay protocol. The glucose
concentrations were 128
± 2 (n = 4), 72.5 ± 0.8 (n = 4), 78.6 ±
0.6, 69.3
± 0.7 mg/dL (n = 4), respectively. Coefficient
of variance < 3%.
PUBLICATIONS
1. Yoon, S.S. and Mekalanos,
J.J. (2006) 2,3-Butanediol synthesis and the
emergence of the
Vibrio
cholerae El Tor biotype. Infection and
Immunity 74 (12): 6547–6556.
2. Schmidt, C. et al (2007).
Regulation of renal glucose transporters during
severe inflammation. Am J Physiol Renal Physiol
292: F804- F811.
3. Jatana, M. (2006)
Inhibition of NF- κB
activation by 5- lipoxygenase inhibitors
protects brain against injury in a rat model of
focal cerebral ischemia. J. Neuroinflammation
3:12. |
|
EnzyChromTM Glucose Assay Kit
(EBGL-100)
Quantitative Colorimetric/Fluorimetric
Glucose Determination
DESCRIPTION
Glucose (C6H12O6) is a key
diagnostic parameter for many metabolic
disorders. Increased glucose levels have been
associated with diabetes mellitus, hyperactivity
of thyroid, pituitary and adrenal glands.
Decreased levels are found in insulin secreting
tumors, myxedema, hypopituitarism and
hypoadrenalism. Simple, direct and
high-throughput assays for measuring glucose
concentrations find wide applications in
research and drug discovery. BioAssay Systems'
glucose assay kit uses a single Working Reagent
that combines the glucose oxidase reaction and
color reaction in one step. The color intensity
of the reaction product at 570nm or
fluorescence intensity at lem/ex = 585/530nm is
directly proportional to glucose concentration
in the sample.
KEY FEATURES
Sensitive and accurate .
Use as little as 20 μL samples. Linear detection
range in 96-well plate: 5 to 300 μM (90 μg/dL to
5.4 mg/dL) glucose for colorimetric assays and 1
to 30 μM for fluorimetric assays.
Simple and high-throughput.
The procedure involves addition of a single
working reagent and incubation for 30 min at
room temperature.
APPLICATIONS:
Direct Assays:
glucose in serum, plasma, urine, saliva, milk,
culture medium and other biological samples.
Drug Discovery/Pharmacology:
effects of drugs on glucose metabolism.
Food and Beverages:
glucose in food, beverages etc.
KIT CONTENTS:
Assay Buffer:
10
mL Enzyme Mix: 120 μL
Dye Reagent: 120 μL
Standard: 1 mL 300 mg/dL Glucose
Storage conditions. The
kit is shipped on dry ice. Store all componets
at -20°C. Shelf life of three months after
receipt.
Precautions: reagents are
for research use only. Normal precautions for
laboratory reagents should be exercised while
using the reagents. Please refer to Material
Safety Data Sheet for detailed information.
COLORIEMTRIC PROCEDURE
Sample treatment:
saliva samples should be centrifuged for 5 min
at 14,000 rpm prior to assay. Milk samples
should be cleared by mixing 100 μL 6N HCl and
600 μL milk. Centrifuge 5 min at 14,000 rpm and
transfer supernatant into a clean tube. Add 170
μL 6N NaOH per mL supernatant. Mix well and
centrifuge again at 14,000 rpm. The supernatant
can be assayed. The dilution factor in this
procedure is n = 1.36. Samples can be
analyzed immediately after collection, or stored
in aliquots at –20 °C. Avoid repeated
freeze-thaw cycles. If particulates are present,
centrifuge sample and use clear supernatant for
assay.
1. Equilibrate all components
to room temperature. During experiment, keep
thawed Enzyme in a refrigerator or on ice.
2. Standards and samples:
for 300 μM standard, mix 15 μL 300 mg/dL
standard with 818 μL dH2O. Dilute standard in
dH2O as follows.
Transfer 20 μL standards and
samples into separate wells.
3. Working Reagent.
For each reaction well, mix 85 μL Assay Buffer,
1 μL Enzyme Mix (vortex briefly before
pipetting), and 1 μL Dye Reagent in a clean
tube. Transfer 80 μL Working Reagent into each
reaction well. Tap plate to mix.
4. Incubate 30 min at room
temperature. Read optical density at 570nm
(550-585nm).
CALCULATION
Subtract blank OD (water, #4)
from the standard OD values and plot the
DOD
against standard concentrations. Determine the
slope and calculate the glucose concentration of
Sample,
FLUORIMETRIC PROCEDURE
For fluorimetric assays, the
linear detection range is 1 to 30
μM
glucose. Mix 10 μL of the standards from
Colorimetric Procedure with 190 μL dH2O to
obtain standards at 30, 18, 9, 0 μM glucose.
Transfer 20 μL standards and 20 μL samples into
separate wells of a black 96-well plate.
Add 80 μL Working Reagent (see Colorimetric
Procedure), tap plate to mix. Incubate 30
min at room temperature and read fluorescence at
lex = 530nm and lem = 585nm. The glucose
concentration of Sample is calculated as
Notes: (1). If the
calculated sample glucose concentration is
higher than 300 μM
in colorimetric assay or 30 μM in fluorimetric
assay, dilute sample in water and repeat the
assay. Multiply result by the dilution factor.
(2). To determine glucose in phenol red culture
medium, dilute both sample and glucose standards
in the same glucose free medium for colorimetric
assay. For fluorimetric assay, prepare standards
in phenol red medium. Dilute sample and
standards 20-fold or more in water.
MATERIALS REQUIRED, BUT NOT
PROVIDED
Pipetting devices, centrifuge
tubes, clear flat-bottom 96-well plates, black
96-well plates and plate reader.
LITERATURE
1. Okuda J, Okuda G. (1969).
A rapid polarographic microdetermination of
glucose with glucose oxidase. Clin Chim Acta.
23(2):365-7.
2. Saifer A, Gerstenfeld S.
(1958). The photometric microdetermination of
blood glucose with glucose oxidase. J Lab Clin
Med. 51(3):448-60.
3. Middleton JE, Griffiths
WJ. (1957). Rapid colorimetric micro-method for
estimating glucose in blood and C. S. F. using
glucose oxidase. Br Med J. 2(5060):1525-7. |
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