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Galactose and Lactose Colorimetric/Fluorometric Assay Kit

Catalog #: K617

In stock

$415.00

Product Details

Cat # +Size K617-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications N/A
Features & Benefits • Simple procedure; takes ~ 1 hour
• Fast and convenient
• Kit provides all necessary buffers and reagents for a accurate assay of galactose and lactose individually in various biological samples
Kit Components • Galactose Assay Buffer
• Galactose Probe (DMSO solution)
• Lactase (Lyophilized)
• Galactose Enzyme Mix (Lyophilized)
• HRP (Lyophilized)
• Galactose Standard (100nmol/µl)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Galactose (C₆H₁₂O₆ FW: 180.16) and lactose (C₁₂H₂₂O₁₁ FW: 342.3) are naturally occurring sugars. Lactose consists of one galactose and one glucose. Some people, particularly infants, lack the enzyme necessary to digest galactose leading to galactose accumulation in blood (Galactosemia) causing enlarged liver, renal failure, cataracts and brain damage. BioVision’s Galactose / Lactose Assay Kit provides a simple, convenient means for direct measurement of galactose and lactose levels in various biological samples (serum, plasma, other body fluids, food, growth media, etc.). To detect Galactose, galactose oxidase specifically oxidizes free galactose, generating a product that reacts with the Galactose Probe to produce color (O.D 570nm) and fluorescence (Ex/Em 535/587). To detect lactose, lactose is hydrolyzed by lactase to generate free galactose which is then measured. Thus, Lactose level = Total Galactose – Free Galactose.


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If samples contain Allolactose will that interfere with the measurements?
The Galactose assay part will work (without hydrolysis by Lactase enzyme). It turns out that the E.coli Lactase we use in our kit will break down both Lactose and Allolactose into glucose and galactose. This will interfere in the lactose measurement using this kit . We recommend substituting the lactase enzyme from our kit with another beta-galactosidase enzyme from a different organism such that it only hydrolyzes Lactose and not Allolactose.
To measure Lactose without any ineterest in measuring Galactose is this kit still recommended?
We recommend using the K624 kit for Lactose measurements in this case.
Are there detergents in the assay buffer to help lyse cells?
There is <0.5% of NP-40 in the assay buffer. Please refer to the MSDS for this information.
Can protein content be used as an internal control for this assay?
Yes, a detergent-compatible BCA assay can be used for protein quantitation to normalize sample amount.
Can this assay be used with bacterial growth medium which is reducing in nature?
The assay might not work at all under strong reducing conditions because the assay reaction step 2 is based on oxidation of galactose.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.