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MAK045

Sigma-Aldrich

HDL and LDL/VLDL Quantitation Kit

sufficient for 100 colorimetric or fluorometric tests

Synonym(e):

HDL and LDL/VLDL Assay Kit

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About This Item

UNSPSC-Code:
12161503
NACRES:
NA.25

Verwendung

sufficient for 100 colorimetric or fluorometric tests

Anwendung(en)

cosmetics
food and beverages

Nachweisverfahren

colorimetric
fluorometric

Relevante Krankheit(en)

cardiovascular diseases

Lagertemp.

−20°C

Allgemeine Beschreibung

Lipoproteins transport the majority of plasma lipids including cholesterol and triglycerides. High-density lipoprotein (HDL), low-density lipoprotein (LDL), and very-low-density lipoprotein (VLDL) are the lipoproteins responsible for the vast majority of cholesterol transport in the blood. High LDL levels and low HDL levels are strongly associated with increased risk of adverse cardiovascular events.

Anwendung

HDL and LDL/VLDL Quantitation Kit has been used to determine the concentrations of HDL and LDL/VLDL in serum samples.[1][2]

Eignung

Suitable for use with plasma and serum samples.

Prinzip

In this kit, serum HDL and LDL/VLDL are first separated and then the cholesterol concentration of each is determined by a coupled enzyme assay, which results in a colorimetric (570 nm)/fluorometric (λex = 535/λem = 587 nm) product, proportional to the cholesterol present. This kit can also be used to determine the concentration of free cholesterol and cholesteryl esters present in a sample.

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Piktogramme

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Signalwort

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H-Sätze

H334

Gefahreneinstufungen

Resp. Sens. 1

Lagerklassenschlüssel

10 - Combustible liquids

Flammpunkt (°F)

188.6 °F - closed cup

Flammpunkt (°C)

87 °C - closed cup

Hier finden Sie alle aktuellen Versionen:

Analysenzertifikate (COA)

Lot/Batch Number
7K25K06130
8D19K06130
7K01K06130
7C25K06130
6M29K06130

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Fragen

1-10 von 11 Fragen  

  1. How is shipping temperature determined? And how is it related to the product storage temperature?

    1 Antwort

    1. Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf

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  2. How can I determine the shelf life / expiration / retest date of this product?

    1 Antwort

    1. If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
      For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
      For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
      For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/449/386/product-dating-information-mk.pdf

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  3. Which reactive oxygen species are detected by the MAK142, MAK143, MAK144, and MAK145 kits?

    1 Antwort

    1. The specified reactive oxygen species targeted by each kit are as follows:

      • MAK142: Hydrogen Peroxide (H2O2), Hydroxyl radical (•OH), Tert-butyl-hydroperoxide (TBHP), Hypochlorous acid (HOCl), Superoxide anion (O2•-)
      • MAK143: Hydrogen Peroxide (H2O2), Hydroxyl radical (•OH), Tert-butyl-hydroperoxide (TBHP), Hypochlorous acid (HOCl)
      • MAK145: Hydrogen Peroxide (H2O2), Hydroxyl radical (•OH), Tert-butyl-hydroperoxide (TBHP)
      • MAK144: Hydrogen Peroxide (H2O2), Hydroxyl radical (•OH), Tert-butyl-hydroperoxide (TBHP), Superoxide anion (O2•-)
      The principle of the reactive oxygen species (ROS) dyes involves non-conjugated compounds that are concealed with a ROS-cleavable blocking group. Upon the reaction with ROS, the blocking group is removed, releasing a highly fluorescent dye. The ROS production is directly proportional to the concentration of the fluorophore. Additionally, the MAK045 is a cholesterol oxidase based assay kit that recognizes free cholesterol, producing a product that reacts with a probe to generate color.

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  4. Is it best to dilute the serum with PBS prior to the separation, or dilute when adding to the wells? Do you have a guide value for the volume of the HDL or LDL sample that should be added to each well?

    1 Antwort

    1. Serum should be mixed directly with the 2X precipitation buffer followed by incubation and centrifugation at the described times to isolate the HDL supernatant fraction. The precipitant contains the LDL/VLDL fraction and is reconstituted in PBS. Sample volumes between 2-50 uL can be used with adjustment to 50 uL/well with assay buffer.

      Hilfreich?

  5. Does the MAK045 HDL and LDL/VLDL Quantitation Kit have the capability to detect oxidized LDL? Moreover, does the Cholesterol Esterase in the kit recognize oxidized LDL as a substrate?

    1 Antwort

    1. As per information obtained from a quick literature search, it appears that the cholesteryl ester core of oxidized LDL can be influenced by cholesterol esterase. However, no specific tests have been conducted to validate this.

      Hilfreich?

  6. Is the MAK045 HDL and LDL/VLDL Quantitation Kit compatible with blood taken using serum separator tubes (also known as marble-top tubes)?

    1 Antwort

    1. The MAK045 HDL and LDL/VLDL Quantitation Kit is expected to function well with serum collected from a serum separator tube (SST) as well as with serum collected without these tubes.

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  7. Is the reason there is an option for colorimetric and fluorometric for colorimetric to detect higher levels vs fluorometric for lower HDL/LDL levels? Which would you recommend for 8 week old mouse serum?

    1 Antwort

    1. A fluorometric approach is 10X more sensitive than that of a colorimetric assay. This is the reason the standard curve is prepared at a lower concentration range. For determining HDL and LDL/VLDL in serum isolated from 8-week old mice, the following article may be helpful:
      Moody J, Yang C, Sedinkin J, Chang Y. Systemic MCPIP1 deficiency in mice impairs lipid homeostasis. Curr Res Pharmacol Drug Discov. 2020 May 6;1:1-9. doi: 10.1016/j.crphar.2020.03.001. PMID: 34909637; PMCID: PMC8663940.

      Note that the preceding article measured values at 560 nm. It is recommended to perform the assay at 570 nm as indicated in the technical bulletin. Any modifications would have to be validated by the end user.

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  8. Can these kits be used with cell samples or medium samples from hepatic cells?

    1 Antwort

    1. Unfortunately, this kit has been qualified for use with plasma and serum samples only. The suitability of this kit with cell or medium samples has not been validated. This kit has been used successfully with liver samples by other researchers. These publications are listed below:
      Chang et al. 2019
      Yu et al. 2018
      Sadh et al. 2017

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  9. Can these kits be used with serum samples from mouse?

    1 Antwort

    1. Yes, this kit can be used on mouse serum samples. We generally recommend that fresh serum samples be used. Frozen samples can probably be used. However, the successful use of frozen samples depends on the efficiency of freezing them and the amount of time they have been frozen. If previously frozen serum samples are used, you may want to check the performance of the kit with fresh vs. frozen samples.

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  10. For Product MAK045, HDL and LDL/VLDL Quantitation Kit, instead of using microcentrifuge tubes in the sample preparation, have you developed a method that uses a 96-well filter plate?

    1 Antwort

    1. For product MAK045, HDL and LDL/VLDL Quantitation Kit, we have not used a filter plate. The common problem with using filters is that lipids tend to bind to or get stuck in them. Using a filter plate would probably give decent results for the HDL solution that would go through the filter. However, it would be very difficult to get the LDL/VLDL precipitate resuspended completely if it is on a filter membrane in a 96-well plate.

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