SILu™Prot APOA1, Apolipoprotein A-1 human
SIGMA/MSST0001 - recombinant, expressed in HEK 293 cells, SIL MS Protein Standard, 13C- and 15N-labeled
Synonym: SILu™Prot Apolipoprotein A-1
Product Type: Chemical
SILuProt products are stable isotope-labeled full length proteins designed to be used as mass spectrometry internal standards for quantitative proteomics. Introduction of an internal SILuProt standard at the beginning of the MS workflow minimizes variations in quantification due to protein extraction, fractionation, enrichment, proteolysis and analysis
RP-LC-MS Analysis Analysis of intact SILuProt APOA1 (MSST0001) using Supelco BIOshell A400 Protein C4, 150 Ò 2.1 mm, 3.4 µm (66826-U) . The deconvoluted mass spectrum reveals a mixture of the Proapolipoprotein A-I (APOA1 19-267 + HisTag) and fully mature protein (APOA1 25-267 + HisTag). There is no signal sequence in either mass.
SDS-PAGE SDS-PAGE of SILuProt (“heavy”) APOA1 incorporating 13C615N4Arg/13C615N2Lys
Stable Isotope Incorporation Incorporation of stable isotope-labeled amino acids 13C615N4Arg and 13C615N2Lys for representative peptides from SILuProt APOA1 was >99%.
Linearity A. XIC chromatogram of three APOA1 peptides. B. Calibration curves of three SILuProt APOA1 peptides obtained by spiking human serum samples with SILuProt APOA1 at levels from 2 µg/mL to 500 µg/mL.
| assay | ≥98% (SDS-PAGE) |
| biological source | human |
| form | lyophilized powder |
| packaging | vial of ≥10 μg (Lot-specific vial content given on certificate of analysis) |
| Quality Level | 200  |
| recombinant | expressed in HEK 293 cells |
| storage temp. | −20°C |
| tag | His tagged |
| V5 tagged | |
| technique(s) | mass spectrometry (MS): suitable |
| UniProt accession no. | P02647  |
| Analysis Note: | General description SILu™Prot ApoA−Ι is a recombinant, stable isotope-labeled human Apo A1 which incorporates [13C6, 15N4]-Arginine and [13C6, 15N2]-Lysine. Expressed in human 293 cells, it is designed to be used as an internal standard for bioanalysis of ApoA1 in mass-spectrometry. SILu Prot Apo A-Ι is a monomer of 280 amino acids (including C-terminal polyhistidine and V5 tags), with an apparent molecular weight of 32.3 kDa. Sequence RHFWQQDEPPQSPWDRVKDLATVYVDVLKDS LLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQ QPYLDDFQKKWQEEMELYRQKVEPL RDRARAHVDALRTHLAPYSDELRQR AKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQSDPSRGPFEGK PIPNPLLGLDSTRTGHHHHHHHHGG The N-terminal signal peptide and C-terminal linker peptide, V5 and polyhistidine tags are italicized. Suggested transitions for three peptides (bolded) are used for selected reaction monitoring analysis (SRM). Label Incorporation ≥ 98% as determined by mass spectrometry Other Characterization • Sequence confirmed by intact mass analysis • Identity verified by peptide mapping • Purity >98% by SDS-PAGE • Vial content was determined by the Bradford method using BSA as a calibrator. The correction factor of Bradford-to-AAA is 88.33% Suggested Quantitative Analysis Parameters (MRM settings provided for three suggested peptides ) |
| Application: | Posters: Characterization of Heavy Recombinant Proteins for Use as Internal Standards in Quantitative MS Workflows Characterization of stable isotope labeled APO-A1 for use as an internal standard in a quantitative MS workflow Characterization of Clinically-Relevant Stable Isotope Labeled Recombinant Proteins for Use as Internal Standards in Quantitative MS Workflows |
| General description: | ApoA-1 (apolipoprotein A-1) is a 29.0kDa protein produced in the liver and intestine, and secreted as the predominant constituent of nascent high-density lipoprotein (HDL) particle. BP- ApoA-1 (apolipoprotein A-1), which is found exclusively in HDL (high-density lipoprotein), has a unique ability to capture and solubilize free cholesterol. This apoA-1 ability enables HDL to remove excess peripheral cholesterol and return it to the liver for recycling and excretion. The therapeutic potential of apoA-1 has been recently assessed in patients with acute coronary syndromes, using a recombinant form of a naturally occurring variant of apoA-1 (called apoA-1 Milano). The availability of recombinant normal apoA-1 should facilitate further investigation into the potential usefulness of apoA-1 in preventing atherosclerotic vascular diseases. Changes in the level of serum apoA-1 may serve as a prognostic marker for non-metastatic nasopharyngeal carcinoma. Low levels of apoA-1 in the plasma are linked to hyperhomocysteinemia. |
| Legal Information: | SILu is a trademark of Sigma-Aldrich Co. LLC |
| Legal Information: | This product is licensed under U.S. Patent No. 7,396,688 and foreign counterparts from E. I. du Pont de Nemours and Company. The purchase of this product conveys to the buyer the nontransferable right to use the purchased amount of the product for research and development only, including services for a third party for consideration. The buyer cannot sell or otherwise transfer this product, its components or materials made using this product or its components to a third party. Information about licenses for excluded uses is available from: E. I. du Pont de Nemours and Company; Attn: Associate Director, Commercial Development; DuPont Experimental Station E268; 200 Powdermill Rd.; Wilmington, DE 19803; 1-877-881-9787 (voice), 1-302-695-1437 (fax), licensing@dupont.com. |
| Physical form: | Supplied as a lyophilized powder containing phosphate buffered saline. |
| Preparation Note: | Briefly centrifuge the vial before opening. It is recommended to reconstitute the protein in sterile ultrapure water to a final concentration of 100μg/ml. |
| Preparation Note: | Store the lyophilized product at –20 oC. The product is stable for at least 2 years as supplied. After reconstitution, it is recommended to store the protein in working aliquots at –20 oC. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Purity | ≥98% (SDS-PAGE) |
| Storage Temp. | −20°C |
| UNSPSC | 23201100 |