![Western Blot: Raw 264.7 Whole Cell Lysate [NBP1-30157] - Samples: 25ug whole cell lysate of RAW 264.7 cells treated with T0901317 (lane 1) and untreated (lane 2) [NBP1-30157].](http://images.novusbio.com/fullsize/Raw-264.7-Whole-Cell-Lysate-Western-Blot-NBP1-30157-img0003.jpg "Western Blot: Raw 264.7 Whole Cell Lysate [NBP1-30157] - Samples: 25ug whole cell lysate of RAW 264.7 cells treated with T0901317 (lane 1) and untreated (lane 2) [NBP1-30157].")
| Reactivity | MuSpecies Glossary |
| Applications | WB |
| Concentration | 1.0 mg/ml |
| Dilutions |
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| Application Notes | This lysate is prepared as a positive control for separation by SDS-PAGE and subsequent Western blot analysis. Lysates are prepared in denaturing buffer WITHOUT reducing agents (i.e. 2-mercaptoethanol (BME) or dithiothreitol (DTT)). If reducing conditions are desired, add a reducing agent prior to heating. Heat lysate to 95C for 5 minutes and rapidly cool. NOTE: Boiling can cause aggregation in large proteins, resulting in the proteins inability to enter the gel. Large protein samples should be heated to 65C for 5-10 minutes prior to loading. |
| Control |
| Storage | Store at -80C. Avoid freeze-thaw cycles. |
| Buffer | Standard RIPA buffer including protease and phosphatase inhibitors and 0.1% SDS diluted in 1X lithium dodecyl sulfate (LDS) sample buffer with bromophenol blue and 40-70% glycerol, pH 8.4. BME has not been added. |
| Concentration | 1.0 mg/ml |
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![Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Basal ABCA1 expression & function in patient-derived iRPEs. (A) Representation of patient-derived iPSC lines & genotypes harboring polymorphisms in ABCA1 that are associated with decreased or increased risk for AMD development. (B) Relative expression of ABCA1 mRNA in patient-derived iRPEs under basal conditions normalized to RPL28 & decreased risk group. (C) Western blot analysis of ABCA1 levels in patient-derived iRPEs under basal conditions. Actin was detected as loading control. ABCA1 expression was quantified & normalized to actin. (D,E) Relative expression of ABCG1 (D) & NR1H3 (E) mRNA in patient-derived iRPEs under basal conditions normalized to RPL28 & decreased risk group. (F) Cholesterol efflux in patient-derived iRPEs after direct cell labeling & in the presence of ApoAI. (G) Cholesterol efflux in patient-derived iRPEs after phagocytosis of BODIPY-cholesterol-loaded POSs & in the presence of ApoAI. (H) Quantification of phagocytosed POSs per nuclei in patient-derived iRPEs. Values shown are means ± SD (n = 3). * p < 0.05; *** p < 0.001. Unpaired Student’s t-test. (I) Representative fluorescence microscopy images of phagocytosed FITC-labeled POSs (green) & staining for ZO-1 (red) & DAPI (blue) of increased risk cell line IPS19-00096. Scale bar = 100 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35328615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](https://images.novusbio.com/images/nb400-105_rabbit-polyclonal-abca1-antibody-310202416171459.jpg "Western Blot: ABCA1 Antibody - BSA Free [NB400-105] - Basal ABCA1 expression & function in patient-derived iRPEs. (A) Representation of patient-derived iPSC lines & genotypes harboring polymorphisms in ABCA1 that are associated with decreased or increased risk for AMD development. (B) Relative expression of ABCA1 mRNA in patient-derived iRPEs under basal conditions normalized to RPL28 & decreased risk group. (C) Western blot analysis of ABCA1 levels in patient-derived iRPEs under basal conditions. Actin was detected as loading control. ABCA1 expression was quantified & normalized to actin. (D,E) Relative expression of ABCG1 (D) & NR1H3 (E) mRNA in patient-derived iRPEs under basal conditions normalized to RPL28 & decreased risk group. (F) Cholesterol efflux in patient-derived iRPEs after direct cell labeling & in the presence of ApoAI. (G) Cholesterol efflux in patient-derived iRPEs after phagocytosis of BODIPY-cholesterol-loaded POSs & in the presence of ApoAI. (H) Quantification of phagocytosed POSs per nuclei in patient-derived iRPEs. Values shown are means ± SD (n = 3). * p < 0.05; *** p < 0.001. Unpaired Student’s t-test. (I) Representative fluorescence microscopy images of phagocytosed FITC-labeled POSs (green) & staining for ZO-1 (red) & DAPI (blue) of increased risk cell line IPS19-00096. Scale bar = 100 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35328615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")