Anti-LRP1/Lrp 1 Cluster Ii Rabbit Monoclonal Antibody

Western blot analysis of LRP1 expression in A549 cell lysate.

Western blot analysis of LRP1/Lrp 1 Cluster Ii using anti-LRP1/Lrp 1 Cluster Ii antibody (M00829).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human U87 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human U20S whole cell lysates,
Lane 4: human Hela whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat heart muscle tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-LRP1/Lrp 1 Cluster Ii antigen affinity purified monoclonal antibody (M00829) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:1000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for LRP1/Lrp 1 Cluster Ii at approximately 85 kDa. The expected band size for LRP1/Lrp 1 Cluster Ii is at 85 kDa.

Membrane-receptor LRP1 is responsible for the stimulatory effect of CD44s-tPA on lamellipodia formation (A) The distribution patterns of cortactin (red) and F-actin (green) demonstrated by immunofluorescence staining in control and LRP1 inhibitor (RAP, 200 nM), Annexin A2 inhibitor (LCKLSL, 2.5 µM), and EGFR inhibitor (AG1478, 10 µM) pretreated CD44s-overexpressing MCF7 cells. The elongated lamellipodia are highlighted by the arrows. The proportion of cells with lamellipodia in control and RAP, LCKLSL, and AG1478 pretreated groups were calculated from triplicate independent experiments, means ± SD from triplicate experiments were plotted. (B) Representative images and quantitative analysis of migration assay showing the wound closure rate of control and RAP pretreated MCF7 CD44s cells. The means ± SD of wound closure rates from triplicate experiments were plotted. (C) Analysis of LRP1 expression in control and LRP1 knockdown MCF7CD44s cells. (D) Representative images and quantitative analysis of migration assay showing the wound closure rate of control and LRP1 knockdown MCF7 CD44s cells. The means ± SD of wound closure rates from triplicate experiments were plotted. (E) Representative images and quantitative analysis of migration assay showing the wound closure rate of control and LRP1 knockdown MCF7 CD44s cells. The means ± SD of wound closure rates from triplicate experiments were plotted. n. s. Indicates no significant, ** p < 0.01, *** p < 0.001.
Index in PubMed under a CC BY license. PMID: 37842093

tPA/LRP1 axis enhances the lamellipodia formation through NFκB signaling pathway. (A) Gene set enrichment analysis (GSEA) enrichment plots of the hallmark of NFκB gene sets in MCF7 CD44s compared with MCF7 vector groups. (B) Analysis of phosphorylation of p65, total p65 protein levels in MCF7 vector , MCF7 CD44s , and LRP1 knockdown MCF7 CD44s cells by western blot. (C) Analysis of phosphorylation of p65, total p65 protein levels in control and RAP pretreated MCF7 CD44s cells by western blot. (D) Analysis of phosphorylation of p65, total p65 protein levels in control and NFκB pathway inhibitor (BAY 11-7082, 10 µM) pretreated MCF7 CD44s cells by western blot. (E) The distribution patterns of cortactin (red) and F-actin (green) demonstrated by immunofluorescence staining in control and BAY 11-7082 pretreated CD44s-overexpressing MCF7 cells. The elongated lamellipodia are highlighted by the arrows. The proportion of cells with lamellipodia in control and BAY 11-7082 pretreated groups were calculated from triplicate independent experiments, means ± SD from triplicate experiments were plotted. (F) Representative images and quantitative analysis of migration assay showing the wound closure rate of control and BAY 11-7082 pretreated MCF7 CD44s cells. The means ± SD of wound closure rates from triplicate experiments were plotted. * p < 0.05.
Index in PubMed under a CC BY license. PMID: 37842093

All lanes use the Antibody at 1:3W dilution for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human liver carcinoma, using LRP1 Antibody.

Immunohistochemical analysis of paraffin-embedded Human pituitary tumor, using the Antibody at 1:300 dilution.

Immunohistochemical analysis of paraffin-embedded Human prostate cancer, using the Antibody at 1:300 dilution.

Immunohistochemical analysis of paraffin-embedded Human breast cancer, using the Antibody at 1:300 dilution.

Immunofluorescent analysis using the Antibody at 1:50 dilution.

Flow cytometric analysis of LRP1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).