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References

This product has been referenced in:

  • 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 . Read more (PubMed: 23251382) »
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Customer reviews and Q&As

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This product does not contain phosphotungstic acid.

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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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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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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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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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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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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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