HDL and LDL/VLDL Cholesterol Assay试剂盒(ab65390)

概述

  • 产品名称HDL and LDL/VLDL Cholesterol Assay试剂盒
    参阅全部 HDL LDL/VLDL 试剂盒
  • 测试
    100 x 1 assay
  • 样品类型
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids
  • 检测类型Quantitative
  • 检测时间
    1h 00m
  • 种属反应性
    与反应: Mouse, Rat, Rabbit, Human
    预测可用于: all Mammals
  • 产品概述

    Abcam's HDL and LDL/VLDL Cholesterol Assay Kit (ab65390) provides a simple quantification method of HDL and LDL/VLDL after a convenient separation of HDL from LDL and VLDL (very low-density lipoprotein) in serum samples. In this assay, cholesterol oxidase specifically recognizes free cholesterol and produces a component that will react with probe to generate color (570 nm) and fluorescence (Ex/Em = 538/587 nm). Cholesterol esterase hydrolizes cholesteryl ester into free cholesterol, therefore, cholesterol ester and free cholesterol can be detected separately in the presence and absence of cholesterol esterase in the reactions.



    Visit our FAQs page for tips and troubleshooting.

  • 说明

    Regulation of HDL (high-density-lipoprotein)-cholesterol and LDL (low-density-lipoprotein)-cholesterol plays a central role in various disease developments. It is well known that low levels of HDL and high level of LDL are associated with an increased risk of cardiovascular events.

     

    If you need to purchase additional HDL precipitation buffer, please see ab105138.

  • 经测试应用适用于: Functional Studiesmore details

性能

  • 相关性HDL (high-density-lipoprotein), LDL (low-density lipoprotein) and VLDL (very low-density lipoprotein) are lipoproteins that act as carrier proteins for cholesterol. The regulation of HDL-cholesterol and LDL-cholesterol plays a central role in various disease development. It is well known that low levels of HDL and high level of LDL are associated with an increased risk of cardiovascular events.
  • 别名
    • High density lipoprotein
    • LDL
    • Low density lipoprotein
    • Very low density lipoprotein
    • VLDL
    see all

应用

Our Abpromise guarantee covers the use of ab65390 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Functional Studies Use at an assay dependent dilution.

HDL and LDL/VLDL Cholesterol Assay Kit 图像

  • Rat serum samples were processed according to the protocol. The quantity of total (dilution range 1:10-1:100), HDL (dilution range 1:1-1:10), LDL/VLDL (dilution range 1:1-1:10) and free cholesterol (neat) was measure colourimetrically, in duplicates, after 40 minutes.

  • Signal from standard curve was measured colourimetrically over a period of time. Background signal subtracted and each point on the curve represents duplicate values (+/- SD).

  • Measurement of total cholesterol, HDL, LDL/VLDL from serum samples. Total Cholesterol (blue), HDL (green), and LDL/VLDL (cream) cholesterol were measured following the kit protocol.
  • Wild type (WT) or farnesoid X receptor knock-out (FRX-KO) mice were treated with either vehicle (Veh) or a high fat-containing diet (HFD) for 16 weeks. Serum HDL (top) and LDL/VLDL cholesterol (bottom) from 5-7 mice/group was measured using ab65390 following protocol instructions. An asterisk (*) means P<0.05 between WT and FXR-KO vehicle group. A pound (#) indicates P<0.05 between Wt and FXR-KO-HFD group. Image obtained from Li G et al., PLoS One, 2012; 7(4): e35895

实验方案

HDL and LDL/VLDL Cholesterol Assay Kit (ab65390)参考文献

This product has been referenced in:
  • Grasa-López A  et al. Undaria pinnatifida and Fucoxanthin Ameliorate Lipogenesis and Markers of Both Inflammation and Cardiovascular Dysfunction in an Animal Model of Diet-Induced Obesity. Mar Drugs 14:N/A (2016). Rat . Read more (PubMed: 27527189) »
  • Cui XB  et al. RGC32 deficiency protects against high-fat diet-induced obesity and insulin resistance in mice. J Endocrinol 224:127-37 (2015). Functional Studies ; Mouse . Read more (PubMed: 25385871) »

See all 14 Publications for this product

Product Wall

Abreviews
TISSUE PREPARATION
For total cholesterol
1. Homogenize 10 mg tissue in 200 μl of chloroform:Isopropanol:NP-40 (7:11:0.1) using a micro-homogenizer.
2. Spin the extract 10 minutes at 15,000 x g at RT in a centrifuge.
3. Transfer all of the liquid (organic phase) avoiding the pellet, to a new tube. Air dry the sup at 50°C to remove chloroform for 7min (in a hood).
4. Dissolve dried sup with Cholesterol Assay Buffer (1:1) by vortexing until homogeneous (it is OK if the solution becomes cloudy).

Note: The extraction procedure can be scaled up if larger amounts of sample are desired.

For the separation of HDL and LDL/VLDL
1. Use 0.5% NP-40 in water to homogenize the tissue and then combine with the 2X precipitations buffer.
2. Mix 100 µL of sample with 100 µL of 2X Precipitation Buffer (1:1) in microcentrifuge tubes.
3. Incubate 10 minutes at room temperature.
4. Centrifuge at 2,000 x g (5,000 rpm on a bench-top microcentrifuge) for 10 minutes.
5. Transfer the supernatant into new labeled tubes. This is the HDL fraction.
6. Precipitates contain the LDL/VLDL fraction. To measure the LDL/VLDL fraction, centrifuge the precipitate again at 2,000 x g (5,000 rpm on a bench-top microcentrifuge at RT) for 10 minutes and remove trace amount of HDL supernatant carefully. I tissue homogenate you cannot measure the LDL fraction, only HDL!!!! (In tissue culture it is possibale).
7. Resuspend the precipitate in 200 µL PBS. This is the LDL/VLDL fraction.

If the supernatant is cloudy, the sample should be re-centrifuged. If the sample remains cloudy, dilute the sample 1:1 with PBS and repeat the separation procedure from step 2.
Username

Abcam user community

Verified customer

提交于 Mar 29 2016

Abcam HDL/VDL/Total cholesterol

Excellent Excellent 5/5 (Ease of Use)
Abreviews
I had no problems using this kit, it worked good on extracted lipids from liver tissue and values comparable to plasma levels.
Main limitation is the small amount of cholesterol assay buffer provided which just about fitted 100 samples. But we were aware of this and planned accordingly.
We extracted lipids from liver using one of the FAQ questions on the Abcam website related to ab65390 and that seemed to work.
All in all the experiment went good, we opted to assay total cholesterol and HDL only in plasma and liver samples which were 6 months old.
Username

Abcam user community

Verified customer

提交于 Jul 09 2015


I can confirm that this kit is not antibody-based. A chemical polymer is used.

We aim to provide as much information as possible to customers, however I am sorry that on this occasion the exact details are proprietary.

I have contacted the lab regarding your question for ab65390.

After mixing with the Precipitation buffer and centrifuging, there should indeed be a pellet which would be the LDL fraction. The LDL fraction could be sticking to the walls of the ...

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ab65390 HDL and LDL/VLDL Cholesterol Assay Kit can detect 1 ug and 0.1 ug cholesterol by the colorimetric and fluorometric versions respectively. Inter-assay variability from in-house QC results are within 5-10 %. Intra-assay variability is ˜2-3...

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Unfortunately, this kit can only be read on a colorimeter at an OD of 570 nm and on a fluorometer at Ex/Em = 538/587 nm.



Thank you for your inquiry.

A couple of points to note:

- Please subtract the zero std from all std values before making the std curve.

- Our kit directions mention preparing std from 0. 0.1, 0.2, 0.3, 0.4 ...

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EDTA is not ideal for measurement of HDL/LDL in samples. EDTA plasma has potential disadvantages that have discouraged its use. Inadequate mixing can result in microclots. Also, EDTA osmotically draws water from red cells, diluting the plasma constitue...

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You can store the plasma/serum at -80°C in aliquots before assaying with this kit.

Thank you for your enquiry.

Frozen serum when thawed after a considerable time sometimes does show some ppt, but that generally dissipates once the serum is completely thawed and mixed thoroughly. This phenomenon commonly occurs when one thaw...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"