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Lactose Colorimetric/Fluorometric Assay Kit

based on 1 citations in multiple journalsLactose Colorimetric/Fluorometric Assay Kit14.1 4
Sensitive Assay, HTS
Catalog #: K624

In stock

$395.00

Product Details

Cat # +Size K624-100
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 contains all necessary reagents for accurate measurement of Lactose
Kit Components • Lactose Assay Buffer
• Probe (DMSO solution)
• Lactase (Lyophilized)
• Lactose Enzyme Mix (Lyophilized)
• HRP (Lyophilized)
• Lactose Standard (100 nmol/µl)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Lactose (C₁₂H₂₂O₁₁ FW: 342.3) is an important naturally occurred disaccharide, consisting of one galactose and one glucose. Milk contains ~2-8% lactose. 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. In the BioVision Lactose Assay Kit, Lactose is hydrolyzed to glucose and galactose. The galactose is subsequently oxidized generating color (O.D 570 nm) and fluorescence (Ex/Em 535/587 nm). Free galactose can be corrected by a background control in the absence of lactase. The Lactose Assay Kit provides a simple, convenient, and sensitive means for direct measurement of lactose levels in various biological samples (serum, plasma, other body fluids, food, growth media, etc.). Pretreatment of samples is not required. The kit can be used as a high throughput assay.


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The sample is diluted in lactose assay buffer. Would this buffer neutralize the reducing environment of the sample?
The assay buffer is very unlikely to exactly neutralize the reductant in the sample. But this depends on how much sample, reduction potential and concentration of reducing agent and the volume of assay buffer this is diluted in. A bigger concern is that the assay might not work at all under strong reducing conditions because the assay reaction step 2 is based on oxidation of galactose.
What is the difference between Lactose measurements with K617 and this assay?
We recommend using the K624 kit for Lactose measurements if there is no interest in Galactose. K617 is not a direct measurement of lactose. The difference between total and free galactose is calculated as the lactose concentration in K617. This is done to enable measurement of both galactose and lactose in the same sample.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
At what point in the protocol should the Lactase be added?
2ul Lactase should be added to the background control sample before adding the reaction mix. A parallel well with 2ul assay bufer must be prepared for comparison of Glacatose background.
Can this kit be used to assay Lactose in dairy products like milk and yogurt?
Dairy products can be used with this kit. Since they have not been specifically tested, we strongly suggest testing several doses of the sample to make sure the readings are within the standard curve linear range. It is also important to ensure that the samples are not turbid since this can affect absorbance measurements. Free galactose interferes with the assay. Since galactose is present in dairy products, we recommend to prepare two wells for each sample. Add 2 ul of Lactase to one well, add 2 ul of assay buffer to the other well for galactose background control. Galactose background can be subtracted from the lactose assays.
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.
Can the samples prepared with the assay buffer from K617 be used for this assay?
We do not recommend swapping the buffers since optimal conditions (pH, salts etc.) for the specific assay are provided by the assay buffer. In this case, the assay buffer from K617 might work in situations when sample is scarce and there is an absolute need to conserve the samples. We have not tested this when we tested the performance of K624.
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 based on the amount of lactose in the sample. We recommend doing a pilot expt with multiple sample volumes (1-50ul) 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.
How are samples normalized against protein concentration?
A detergent-compatible protein quantitation assay can be used with the supernatants from cell/tissue lysates or with any other liquid sample in the assay buffer.
L L Hernandez, et. el. Suppression of lactation and acceleration of involution in the bovine mammary gland by a selective serotonin reuptake inhibitor. J. Endocrinol., Apr 2011; 209: 45 - 54.
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