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Rabbit anti lrp1

Manufactured by Abcam
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Rabbit anti-LRP1 is an antibody that binds to the LRP1 (low-density lipoprotein receptor-related protein 1) protein. LRP1 is a multifunctional endocytic receptor involved in various cellular processes. This antibody can be used for the detection and analysis of LRP1 in various applications.

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Lab products found in correlation

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3 protocols using rabbit anti lrp1

1

Western Blot Analysis of TTR and LRP1

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The presence of TTR in human serum after depletion, as well as LRP1 total levels in brains and livers was studied by western blot analysis. Protein extract denatured samples (30–50 μg) were separated in SDS-PAGE gels (10% for LRP1 and 15% for TTR). Proteins were then transferred to nitrocellulose membrane (WhatmanTM Ge healthcare – Protan BA 83), using a wet system (Bio-rad Criterion Blotter). The membranes were blocked 1 hour at RT with 5% powered skimmed milk in PBS containing 0,05% Tween-20 (PBS-T). After blocking, membranes were then incubated with primary antibodies in 3% powered skimmed milk/PBS-T against the proteins under study: rabbit anti TTR (DAKO, 1:2000), rabbit anti LRP1 (Abcam, 1:15000), mouse anti Tubulin (Sigma-Aldrich, 1:2000), mouse anti β-Actin (Sigma-Aldrich, 1:3000). Then washed membranes were incubated for 1 hour at RT with sheep anti-rabbit (The binding Site; 1:10000) or anti-mouse (The binding Site; 1:2500) immunoglobulins conjugated with horseradish peroxidase in 3% powered skimmed milk/PBS-T. The blots were developed using ClarityTM Western ECL substrate (Bio-rad) and proteins were detected and visualized using a chemiluminescence detection system (ChemiDoc, Bio-rad).
Alemi M., Gaiteiro C., Ribeiro C.A., Santos L.M., Gomes J.R., Oliveira S.M., Couraud P.O., Weksler B., Romero I., Saraiva M.J, & Cardoso I. (2016). Transthyretin participates in beta-amyloid transport from the brain to the liver- involvement of the low-density lipoprotein receptor-related protein 1?. Scientific Reports, 6, 20164.
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2

Western Blot Analysis of Cell Lysates

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The cells were lysed in the SDS-buffer with protease inhibitors (same as for immunoprecipitation). The protein concentration was defined by the Pierce BCA Protein Assay Kit (Life Technologies). The Western blot analysis was performed according to a standard protocol as outlined in Hennen et al. (2011) (link), the images were acquired using a MicroChemie Chemiluminescence-Reader (Biostep, Burkhardtsdorf, Germany). The following antibodies were used: rabbit anti-LRP1 1:10,000 (Abcam, Cambridge, UK), mouse anti-MAP2 (clone AP20, Millipore, Schwalbach, Germany) 1:2000, rabbit anti-PDGFRα (Santa Cruz Biotechnology) 1:3000, mouse anti-GFAP 1:3000 (Sigma-Aldrich), mouse anti-α-tubulin (clone DMA1, Sigma-Aldrich) 1:10,000, rabbit-anti pERK1/2 Thr202/Tyr204 (Cell signaling, Cambridge, UK 1:1000, rabbit-anti ERK1/2 (Santa Cruz Biotechnology) 1:1000, rabbit-anti pAkt Ser473 (Cell signaling) 1:1000, rabbit-anti Akt (Santa Cruz Biotechnology) 1:1000, mouse anti-actin (BD Bioscience, Erembodegem, Belgium) 1:5000, mouse anti-βIII-tubulin (Sigma-Aldrich) 1:500.
Safina D., Schlitt F., Romeo R., Pflanzner T., Pietrzik C.U., Narayanaswami V., Edenhofer F, & Faissner A. (2016). Low-density lipoprotein receptor-related protein 1 (LRP1) is a novel modulator of radial glia stem cell proliferation, survival and differentiation. Glia, 64(8), 1363-1380.
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3

Immunohistochemical Analysis of LRP1 and LeX

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For the immunohistochemical staining the E12.5 embryos were fixed in 4% (w/v) PFA in PBS ON followed by subsequent incubation in 10%, 20% and 30% (w/v) sucrose (each 12 h), and frozen in Tissue Freezing Medium (Leica, Solms, Germany). 40 μm-thick free-floating frontal sections were cut, blocked with 10% (w/v) normal goat serum (NGS), 0.1% (v/v) Triton in PBS for 1 h. The sections were incubated with primary antibodies: rabbit anti-LRP1 1:500 (Abcam), rat anti-LeX 5750 mAb 5750LeX 1:50 (Hennen et al., 2011 (link)) ON in the blocking solution, washed 3×30 min with PBS and incubated with appropriate secondary antibodies: Alexa Flour 488 (Life Technologies) 1:500, Cy3 (Dianova) 1:500 and Topro3 (BioTechniques, New York, NY, USA) 1:500 for 2 h, washed again 3×30 min with PBS and mounted with Immumount (ThermoScientific).
Safina D., Schlitt F., Romeo R., Pflanzner T., Pietrzik C.U., Narayanaswami V., Edenhofer F, & Faissner A. (2016). Low-density lipoprotein receptor-related protein 1 (LRP1) is a novel modulator of radial glia stem cell proliferation, survival and differentiation. Glia, 64(8), 1363-1380.
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