We investigated the expression of LRP1 in the hearts of 6 patients with ischemic heart failure undergoing heart transplant using western blot, and 6 subjects who had died of traumatic death and free of cardiac disease. A portion of the left ventricle, close to the apex, was taken and snap frozen with liquid nitrogen and then stored at –80°C. The samples were then thawed and processed for western blot using a rabbit monoclonal LRP1 antibody (Abcam, ab-92544 Cambridge, United Kingdom). We then studied LRP1 expression at western blot and immunofluorescence in the heart of 8 mice that had had myocardial ischemia surgery 24 h previously and 4 mice that underwent sham surgery. The western blot was performed using a similar approach as outlined for the human samples using glyceraldehyde 3-phosphate dehydrogenase as loading control protein. Immunofluorescence was performed on tissue slides derived from formalin-fixed paraffin-embedded myocardial samples, using the anti-LRP1 antibody (Abcam, ab-92544) and anticardiac myosin heavy chain (Sigma-Aldrich) to stain for cardiomyocytes.
Ab 92544
Manufactured by Merck Group
Ab-92544 is a laboratory equipment product designed for general scientific applications. Its core function is to facilitate the analysis and processing of samples in a controlled and efficient manner. The specific details and intended use of this product are not available.
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2 protocols using ab 92544
1
LRP1 Expression in Ischemic Heart Failure
2
Western Blot Analysis of Alzheimer's Markers
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Cerebral cortex extracts were loaded onto 10–16% Tris/tricine SDS gels and transferred to nitrocellulose membranes before overnight incubation with one of the following primary antibodies: mouse anti-β-site amyloid precursor protein-cleaving enzyme 1 (BACE1) (1:1000; Millipore, #MAB5308), anti-PS1 (1:1000; Sigma-Aldrich, #PRS4203), anti-neprilysin (NEP) (1:1000; Millipore, #AB4348), anti-insulin-degrading enzyme (IDE) (1:1000; Abcam, #ab32216), anti-low-density lipoprotein receptor-related protein (LRP1) (1:1000; Abcam, #ab92544), anti-APP (C-terminal) (1:1000; Sigma-Aldrich, #A8717), mouse anti-total Aβ (Aβ1–16) (1:1000; Covance, #803001), and anti-apoE (1:1000; Abcam, #ab20874) or GAPDH (1:1000; Bioworld; #AP0063). Horseradish peroxidase-conjugated secondary antibodies (Vector Laboratories) were used, and bands were visualized using ECL plus detection system. GAPDH was utilized as an internal control for protein loading and transfer efficiency.
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