The sample volume required for Kit MAK309 is optimized as per the specifications provided in the kit bulletin. Diluting the plasma 20X is not advisable as it would result in a loss of assay sensitivity. Upon confirming with the vendor, it was advised that the sample volume can be scaled down to half the volume in each step, with the other components adjusted accordingly. However, diluting the samples is not recommended as it would compromise sensitivity. Alternatively, the assay can be adapted for use in a 384-well plate or a half-area 96-well plate, such as CLS3880, which have smaller well volumes and can accommodate smaller sample volumes. For further details, refer to the Scaling Down Volume Procedure from the Vendor document.
MAK309
Bile Acid Assay Kit
sufficient for 100 fluorometric tests
Synonym(s):
Biliary Acid Quantification Kit, Total Bile Acids Assay Kit
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1 KIT
$743.00
About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Recommended Products
detection method
fluorometric
relevant disease(s)
gastrointestinal diseases; cancer; cardiovascular diseases
storage temp.
−20°C
Related Categories
General description
Twelve different types of bile acids are typically found in mammals, among them two primary types are cholic acid and chenodeoxycholic acid.[1] These can be dehydroxylated into secondary bile acids. Finally, these four can be conjugated to either taurine or glycine creating 8 different conjugated bile acids. Bile acid levels in feces, blood, urine, and bile can be used as markers for various diseases such as hyperlipidemia, cholestasis, gall stones, colon cancer, etc.[2][3] Bile acids also exist as sulfate salt forms known as bile acid sulfates. Sulfation of bile acids increases their solubility and decreases intestinal absorption, thereby enhancing fecal and urinary excretion.
Application
Bile Acid Assay Kit has been used to quantify bile acid levels from plasma.[4]
Features and Benefits
Compatible with high-throughput handling systems.
Suitability
Suitable for the quantitative determination of total bile acids and evaluation of drug effects on bile acid metabolism in serum, plasma, urine and other biological samples.
Principle
This Bile Acid Assay Kit provides a convenient fluorimetric means to measure total bile acids in biological samples. In the assay, 3–hydroxysteroid dehydrogenase reacts with all twelve bile acids, converting NAD to NADH, which reduces a probe to a highly fluorescent product. The resulting fluorescence intensity (γex = 530 nm/γem = 585 nm) is linear to the bile acid concentration in the sample.
Storage Class Code
10 - Combustible liquids
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Is it feasible to dilute mouse plasma 20X for use with the Bile Acid quantification kit (catalog number: MAK309), considering that it requires a significant amount of samples (20 µl per well, in duplicate)?
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Can bile acids from HepG2 cell culture supernatants be checked using this kit?
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Unfortunately, the kit has not been validated with cell culture supernatant, and there are no relevant citations for this particular use. While the expected level of bile acid in HepG2 cell culture supernatant is unknown, given that significant amounts of unconjugated bile acids are released into the medium of HepG2 cell cultures, the sample might be compatible with the kit. It is advisable to conduct an initial serial dilution of the sample to identify the appropriate dilution factor that yields results within the linear detection range (1-150 uM bile acid). The Sample Blank should be adjusted to accommodate possible interference in the sample matrix. If the initial results are not satisfactory, further dilution of the sample may be necessary.
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Is it possible to use the MAK309 bile acid assay kit to measure unknown quantities of bile acids in a solution, such as NaOH? Would NaOH interfere with the assay, and can standards of known bile acid concentration in the same solution be used to address this?
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The MAK309 assay has not been tested in a NaOH solution. It is believed that it should work, but if the NaOH solution is basic, it may interfere with the enzymes in the kit. Neutralizing the pH of the NaOH solution before performing the assay is recommended if possible. Additionally, a test run of the kit could be conducted by preparing the internal standard in NaOH. It is important to use NaOH as the blank instead of water to adjust for any changes in the measured bile acid concentration due to the use of NaOH.
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Is a solid, black round 96-well bottom plate sufficient for the assay, or is a 96-well black plate with a clear bottom necessary?
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A black, round-bottom plate can be used for this assay instead of a black, clear-bottom plate. Although there may be slight variations in the standard curve (resulting in a higher slope compared to a black flat-bottom plate), the standard curve remains linear, making the black round-bottom plate suitable for use.
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I am looking for the protocol for this bile acids kits MAK309-1KT
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The technical bulletin for the Product MAK309 can be accessed in the DOCUMENTATION section of the Product Detail Page under 'More Documents'. Please see the links below:
https://www.sigmaaldrich.com/product/sigma/mak309#product-documentation
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/350/583/mak309bul.pdfHelpful?
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Hi, I'm looking for a Bile Acid Assay Kit to detect bile acids in stool. Do you have any information/products that may be useful for this purpose? Thanks, Alicia
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Product MAK309, Bile Acid Assay Kit, is a fluorometric assay kit that is suitable for the quantitative determination of total bile acids and evaluation of drug effects on bile acid metabolism in serum, plasma, urine and other biological samples. For assay information, please review this data sheet: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/350/583/mak309bul.pdf
If you would like to search other options , we also offer product MAK382, Total Bile Acids (TBA) Assay kit (Colorimetric), which is a kit that can detect as little as 1μM of bile acids in biological fluids such as serum, plasma, bile, urine, saliva, etc. For assay information, please review this data sheet: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/401/650/mak382bul-ms.pdf
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