MSST0001

SILu^™Prot APOA1, Apolipoprotein A-1 human

recombinant, expressed in HEK 293 cells, SIL MS Protein Standard, ¹³C- and ¹⁵N-labeled

• Synonym(s): SILu^™Prot Apolipoprotein A-1
• UNSPSC Code: 23201100

Properties

• biological source: human
• Quality Level: 200
• recombinant: expressed in HEK 293 cells
• tag: His tagged; V5 tagged
• Assay: ≥98% (SDS-PAGE)
• form: lyophilized powder
• packaging: vial of ≥10 μg (Lot-specific vial content given on certificate of analysis)
• technique(s): mass spectrometry (MS): suitable
• UniProt accession no.: P02647
• storage temp.: −20°C
• Gene Information: human ... APOA1(335)

Description

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.

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

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.

Storage and Stability

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.

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

RHFWQQDEPPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLK
LLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKV
QPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEM
RDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEK
AKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQSDPSRGPFEGK
PIPNPLLGLDSTRTGHHHHHHHHGGQ

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)

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.

SILu is a trademark of Sigma-Aldrich Co. LLC

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable