Tuesday, September 27, 2022

Biotium introduces new line of antibodies to detect exosomes

Figure 1. EC-purified MCF-7-derived exosomes were stained with ExoBrite™ CD9 560/585 FlowC antibody (left).  Specific staining was observed in comparison to the same antibody in the buffer (right).  Exosomes in the PE channel were detected on a Cytoflex LX flow cytometer.  To download the image, please click here
Figure 1. EC-purified MCF-7-derived exosomes were stained with ExoBrite™ CD9 560/585 FlowC antibody (left). Specific staining was observed in comparison to the same antibody in the buffer (right). Exosomes in the PE channel were detected on a Cytoflex LX flow cytometer.

Biotium, a leading supplier and inventor of fluorescent cellular probes for life science research, announced the release of new ExoBrite™ antibody conjugates. These antibodies were specifically curated for optimal detection of the exosome markers CD9, CD63, and CD81 in individual exosome or exosome extracts without any background. ExoBrite™ antibody conjugates are available in formulations for both flow cytometry and western blotting.

Extracellular vesicles (EVs), including exosomes, are increasingly studied for their roles in cell communication and as potential targets for clinical and drug delivery applications. The most common proteins used as exosome markers are CD9, CD63, and CD81, members of the tetraspanin family. While antibodies targeting these proteins are available by commercial suppliers, few are validated or perform well for the detection of EVs or exosomes. ExoBrite™ Flow Antibody Conjugates and ExoBrite™ Western Antibody Conjugates were developed by Biotium scientists for optimal detection of CD9, CD63, and CD81 exosome markers for flow cytometry and western blotting.

The ExoBrite™ Flow Antibody is designed for the detection of isolated or bead-bound EVs by flow cytometry. Antibodies are available in 4 conjugates: ExoBrite™ 410/450 (Pacific Blue™ channel), ExoBrite™ 490/515 (FITC channel), ExoBrite™ 560/585 (PE channel with yellow laser), and ExoBrite™ R-PE ( PE channel). Isotype controls are also available for each conjugate.

The ExoBrite™ Western antibody is designed to detect exosome markers in EV extracts by near-IR fluorescent western blot. Antibodies are available with two near-infrared fluorescent dyes, ExoBrite™ 680/700 and ExoBrite™ 770/800, which provide greater signal-to-noise for western blotting than visible light emitting dyes. An Exobrite™ western antibody to detect calnexin, a membrane protein localized in the endoplasmic reticulum that is not found in exosomes, is also available as a negative control to assess the purity of exosome extracts.

Figure 2. Figure 2. Western detection using Exobrite™ western antibody and MCF-7 cell and exosome lysates, showing tetraspanin protein enrichment and calnexin deficiency in exosomes.  Blots were stained with ExoBrite™ 770/800 Calnexin antibody (exosome negative control) plus (A) ExoBrite™ 680/700 CD9 antibody, (B) ExoBrite™ 680/700 CD63 antibody, or (C) ExoBrite™ 680/700 CD81 antibody Were.  ,  Lane M: Protein marker.  Lane 1: 10 misshapen cell lysate.  Lane 2 : 1 misshapen cell lysate.  Lane 3 : 1 misshapen exosome lysate.  Blots were imaged on a LI-COR Odyssey® Infrared Imaging System.
Figure 2. Western Detection using ExoBrite™ Western Antibodies and MCF-7 Cell and Exosome Lysates Showing Tetraspanin Protein Enrichment and Calnexin Deficiency in Exosomes. Blots were stained with ExoBrite™ 770/800 Calnexin antibody (exosome negative control) plus (A) ExoBrite™ 680/700 CD9 antibody, (B) ExoBrite™ 680/700 CD63 antibody, or (C) ExoBrite™ 680/700 CD81 antibody Were. , Lane M: Protein marker. Lane 1: 10 misshapen cell lysate. Lane 2 : 1 misshapen cell lysate. Lane 3 : 1 misshapen exosome lysate. Blots were imaged on a LI-COR Odyssey® Infrared Imaging System.

The release of ExoBrite™ Antibody Conjugates adds to Biotium’s growing line of customized solutions for exosome and EV research. Biotium also offers ExoBrite™ EV Membrane Staining Kits, which are designed for bright and distinct staining of isolated EVs with little or no background. ExoBrite™ EV Membrane Staining Kits are available with four color options for Pacific Blue™, FITC, PE and APC channels. Learn more about Biotium’s innovative solutions for EV research.

About Biotium

Biotium is a leading life science reagent manufacturer and supplier dedicated to providing high quality and innovative fluorescent instrumentation promoting scientific discovery. Our collaborative team of experienced chemists and biologists, who are at the forefront of fluorescent dye design, apply chemistry-based principles to solve unsolved challenges in the life sciences and medical research. Since its inception in 2001, Biotium has developed over 30 patented technologies that have been licensed to leading life science technology companies around the world. Please visit www.biotium.com for more information.

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