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2X LDL/VLDL Precipitation Buffer (ab105138)

Submit a review Q&A (7)References (2)

Key features and details

  • Assay type: Quantitative

製品の概要

製品の特性

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (2)

ab105138 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab105138 は 2 報の論文で使用されています。

  • Wauquier F  et al. Circulating Human Metabolites Resulting from TOTUM-070 Absorption (a Plant-Based, Polyphenol-Rich Ingredient) Improve Lipid Metabolism in Human Hepatocytes: Lessons from an Original Ex Vivo Clinical Trial. Nutrients 15:N/A (2023). PubMed: 37111121
  • Liu C  et al. Effects of a novel pharmacologic inhibitor of myeloperoxidase in a mouse atherosclerosis model. PLoS One 7:e50767 (2012). Functional Studies ; Mouse . PubMed: 23251382

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1-7 of 7 Abreviews or Q&A

Question



Inquiry: Does thsi contain phosphotungstic acid?

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Answer

Thank you for contacting us.



This product does not contain phosphotungstic acid.

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Question

I am interested in your HDL and LDL/VLDL cholesterol assay kit (ab65390).

I am wondering if your separation method (ab105138) is based on Immunoprecipitation (such as anti-ApoB on LDL) or more classical precipitation methods such as Dextran.

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Answer

The precipitation method used on our HDL and LDL/VLDL Cholesterol Assay Kit (ab65390) is based on the use of a water-soluble non-ionic polymer. The primary advantage of our HDL precipitation buffer (ab105138) over dextran/Mg or other IP buffers is that the constituents are benign. The components in our buffer have no or very little propensity to interact with proteins in the sample solution and can specifically precipitate out desired sized molecules. Our reagents ensure precipitation and concentration beyond the limit of solubility of the precipitants.

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Question

I would like to know whether the HDL and LDL precipitated from plasma by the buffer are intact? I am doing research on the microRNA carried by HDL and LDL， so I am going to isolate the RNA from the purfied HDL and LDL. Do you think whether I can use your reagent?

Thanks for your help.

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Answer

This assay was not designed with this type of downstream application in particular and we have not tested the miRNA and Lipid association after separation of HDL and LDL. It might be possible to isolate RNA but there are RNases which have not been inhibited in our protocol which might chew the miRNA of interest. Some modifications might be necessary to optimize the procedure for this.

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Question

Dear technical team,
Our customer purchased ab105138 and would like to know how to use this buffer.
1. add 100ul of HDL Precipitation buffer to 100ul of plasma
2. vortexing
3. after 20 min, centrifuge for 20 min with 10,000rpm

He is gonig to test following this protocol and letme know this is suitable.
Best regards,

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Answer

Thank you for your enquiry.

This product (ab105138: HDL Precipitation Buffer) isone of the components of ab65390: HDL and LDL/VLDL Cholesterol Assay Kit.



The Protocol can be found on the product datasheet (ab65390) at this site:

https://www.abcam.com/hdl-and-ldl-vldl-cholesterol-assay-kit-ab65390.html



I hope this will be useful for your customer.

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Question

P {margin-top:2px;margin-bottom:2px;} Dear technical team,     Our customer asked about ab105138 HDL precipitation buffer. https://www.abcam.com/HDL-Precipitation-Buffer-ab105138-support.html   Is this isolate only HDL or also HDL3 from serum or plasma sample?   If it is only isolate HDL, is there any product for isolate HDL2, HDL3?   The customer is going to apply HDL to test apolipoprotein ELISA kit.   Check this please.     Best regards, Youngjin     P {margin-top:2px;margin-bottom:2px;} ****************************************************** Technical Support & Marketing Team Product Specialist T: 82-31-790-4356  C: 82-10-8735-5370 DAWINBIO Inc. 601, Blue zone, ITECO. 762, Dukpung-dong, Hanam-city, Gyeonggi-do,Korea T: 82-1588-9741   F: 82-31-790-4360 www.dawinbio.com ******************************************************

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Answer

Thank you for your inquiry. Unfortunately, this buffer has only been tested to precipitate total HDL. If this buffer precipitates and in what efficiency the major subfractions of HDL, namely, HDL2 and HDL3, have not been specifically tested. Unfortunately, we do not have a product to isolate HDL2 and HDL3 separately. Maybe this publication will help your customer: http://www.jlr.org/content/38/11/2353 I am sorry that I do not have a positive answer to your question on this occasion and hope this information will nevertheless be helpful to your customer.

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Question

we use your buffer HDL Precipitation Buffer (ab105138) to isolate HDL and LDL from the same patient. about the purity of the HDL and LDL derived from that buffer and asked details about what kind of buffer this is (is this polyanion or apoB deleting agent?). We assume that you have validated this HDL isolation method before you release it as a commercially available product. Could you please send us your validation data? More specifically:   1)      Any native gel electrophoresis pictures showing the purity of the HDL (supernatant) and the LDL (precipitant) using the buffer 2)      Other methods that you have done to validate purity of the lipoproteins 3)      Info on the type of buffer.   I am not sure if details is proprietary info but we think this is important data if people want to start using this buffer. We believe that once our methods paper is published this would mean that many people will start using this buffer and this will benefit your company so would appreciate your help. Also we would like to acknowledge you in the paper so please indicate how we could do that (for providing this data). Let me know if you have any questions.  

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Answer

Thank you for your patience while I contacted the lab for further information.  They have informed me that the fractions are separated into the HDL and the LDL/VLDL fractions. Hence the LDL and VLDL both will be present in the precipitate. Pease see section IV.1 of data sheet (attached)- “Transfer the supernatant into new labeled tubes. This is the HDL fraction. The precipitates are the LDL/VLDL fraction. If you want to measure the LDL/VLDL level, the precipitate should be spun again, and the trace amount of HDL supernatant carefully removed. Resuspend the precipitate in 200 ml PBS (not provided). This is the LDL/VLDL fraction.” We have not tested the ratio of LDL and VLDL in the precipitated fraction and at this point we do not have any other data. Please let me know if there is anything else I can help you with.    

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Question

 Hello,   We are about to publish a methods paper that measures oxidation of HDL and LDL and we use your buffer HDL Precipitation Buffer (ab105138) to isolate HDL and LDL from the same patient. The reviewer raised concerns about the purity of the HDL and LDL derived from that buffer and asked details about what kind of buffer this is (is this polyanion or apoB deleting agent?). We assume that you have validated this HDL isolation method before you release it as a commercially available product. Could you please send us your validation data? More specifically:   1)      Any native gel electrophoresis pictures showing the purity of the HDL (supernatant) and the LDL (precipitant) using the buffer 2)      Other methods that you have done to validate purity of the lipoproteins 3)      Info on the type of buffer.   I am not sure if details is proprietary info but we think this is important data if people want to start using this buffer. We believe that once our methods paper is published this would mean that many people will start using this buffer and this will benefit your company so would appreciate your help. Also we would like to acknowledge you in the paper so please indicate how we could do that (for providing this data). Let me know if you have any questions.  

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Answer

Thank you for contacting Abcam. The buffers used in the separation of these fractions are very specific to separate the HDL and the LDL fractions. We cannot reveal the composition as it is proprietary and at this point we do not have the GE image of these purified fractions. The data attached from the data sheet shows the number obtained with this kit after fraction separation In addition I hope that the following citations in peer-reviewed journals that we have from use of this kit, will be helpful: Chang, B. et al. Absence of Adipose Differentiation Related Protein Upregulates Hepatic VLDL Secretion, Relives Hepatosteatosis, and Improves Whole Body Insulin Resistance in Leptin-Deficient Mice. J. Lipid Res., 2010; 51: 2132-2142. • Ella, G, et. al. Thrombospondin-4 Regulates Vascular Inflammation and Atherogenesis. . Circ. Res., Nov 2010; 107: 1313 - 1325. • Enns, L. et al. Attenuation of AGE-Related Metabolic Dysfunction in Mice with a Targeted Disruption of the Ca• Subunit of Protein A. J. Gerontol. A. Biol. Sci. Med. Sci. 2009; 64A: 1221-1231. • Frolova, E. et al. Thrombospondin-4 Regulates Vascular Inflammation and Atherogenesis. Circ. Res., Sep 2010; 10.1161/CIRCRESAHA.110.232371. • Kasthuri RS et al. (2007) Mol. Cell. Proteomics 10.1074/mcp.M70023-MCP200. • Mariappan, N. et al. NF-kB-Induced Oxidatve Stess Contributes to Mitochondrial and Cardiac Dysfunction in Type II Diabetes. Cardiovasc. Res. 2010 85: 473-483. • Miller AM et al (2008) J. Exp. Med. 205: 339 - 346.   If there is anything else I can help with, please let me know.  

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