Human Apolipoprotein B/ApoB Antibody

Detection of Human Apolipoprotein B/ApoB by Western Blot. Western blot shows lysates of human plasma. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Apolipoprotein B/ApoB Monoclonal Antibody (Catalog # MAB41242) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Apolipoprotein B/ApoB at approximately 500 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Apolipoprotein B/ApoB in Human Liver. Apolipoprotein B/ApoB was detected in immersion fixed paraffin-embedded sections of human liver using Mouse Anti-Human Apolipoprotein B/ApoB Monoclonal Antibody (Catalog # MAB41242) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in hepatocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human Apolipoprotein B/ApoB by Simple Western^TM. Simple Western shows lysates of human plasma, loaded at 1:50. A specific band was detected for Apolipoprotein B/ApoB at approximately 315 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Apolipoprotein B/ApoB Monoclonal Antibody (Catalog # MAB41242). This experiment was conducted under reducing conditions and using the 66-440kDa separation system.

Detection of Apolipoprotein B/ApoB by Western Blot Particle number increases proportionally with needle-to-processing time. (a) A set of four peripheral blood samples was each collected in n=4 biological replicates Streck Cell-Free DNA BCT. EV enrichment was performed by differential ultracentrifugation (dUC), followed by SEC. EV-enriched fractions were analyzed by NTA, BCA and WB. (b) Particle numbers obtained from the different needle-to-processing times, measured by NTA. Data normalized to the control condition of immediately processed blood. Mean of normalized value is displayed for each needle-to-processing time and statistical significance is assessed using one sample t test (Bonferroni-Holm-adjusted p-values) with *p=0.0175 (24 h), **p=0.0033 (72 h) and **p=0.0033 (1 week). (c) Median size of particles recovered in EV-enriched fractions from the different needle-to-processing times. Mean value is displayed for each needle-to-processing time and statistical significance is assessed using paired t test (Bonferroni-Holm-adjusted p-values) with **p=0.0014 (24 h vs. immediate processing), ***p=0.0006 (72 h vs. immediate processing) and ***p=0.0006 (1 week vs. immediate processing). In (b) and (c) symbols signify independent biological replicates. (d) Representative immunoblotting image to characterize particles obtained according to EV markers, cell-specific markers and apolipoprotein markers. Original blots are presented in Supplementary Fig. S10 online. (e)-(g) Quantification of band intensities in (d). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40624148), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Apolipoprotein B/ApoB by Western Blot Density gradient ultracentrifugation combined with size exclusion chromatography and ultrafiltration recovers high particle numbers and reduces co-isolates. (a) A set of three peripheral blood samples was each collected in n=3 biological replicates in Streck Cell-Free DNA BCT. EVs were isolated using either differential ultracentrifugation (dUC) or density gradient ultracentrifugation (DG), followed by SEC. Following SEC, DG-purified EVs were concentrated either by ultracentrifugation (DG+UC) or by ultrafiltration (DG+UF). EV-enriched fractions were analyzed by nanoparticle tracking analysis (NTA) and western blot (WB). (b) Total particle recovery and (c) median particle size measured by NTA in EV-enriched fractions using different combinations of purification methods. In (b) and (c) mean values are displayed for each processing condition, and in (b) statistical significance is assessed using paired t test (DG+UC vs. DG+UF) with *p=0.0311. In (b) and (c) symbols signify independent biological replicates. (d) Representative immunoblotting of EV-enriched fractions obtained with the different strategies. Original blots are presented in Supplementary Fig. S11 online. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40624148), licensed under a CC-BY license. Not internally tested by R&D Systems.