Biotium introduces new line of antibodies for exosome detection
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Figure 1. EC-purified MCF-7-derived exosomes had been stained with ExoBrite™ CD9 560/585 Flow Antibody (left). Specific staining was seen, in contrast with the identical antibody in buffer (proper). Exosomes had been detected on a CytoFLEX LX move cytometer within the PE channel. 

Biotium, a number one provider and inventor of fluorescent mobile probes for all times science analysis, publicizes the discharge of recent ExoBrite™ Antibody Conjugates. These antibodies had been curated particularly for optimum detection of exosome markers CD9, CD63, and CD81 in remoted exosomes or exosome extracts with little to no background. ExoBrite™ Antibody Conjugates can be found in formulations for each move cytometry and western blotting. 

Extracellular vesicles (EVs), together with exosomes, are more and more studied for his or her roles in cell communication, and as potential targets for diagnostic and drug supply functions. The most typical proteins used as exosome markers are CD9, CD63, and CD81, members of the tetraspanin household. While antibodies focusing on these proteins can be found by industrial suppliers, few are validated or carry out properly for detection of EVs or exosomes. ExoBrite™ Flow Antibody Conjugates and ExoBrite™ Western Antibody Conjugates had been developed by Biotium scientists for optimum detection of CD9, CD63, and CD81 exosome markers for move cytometry and western blotting.

ExoBrite™ Flow Antibodies are designed for detection of remoted or bead-bound EVs by move cytometry. The antibodies can be found 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 additionally obtainable for every conjugate.

ExoBrite™ Western Antibodies are designed for detection of exosome markers in EV extracts by near-IR fluorescent western blot. The antibodies can be found with two near-infrared fluorescent dyes, ExoBrite™ 680/700 and ExoBrite™ 770/800, which supply higher signal-to-noise for western blotting than dyes with seen mild emission. An ExoBrite™ Western Antibody for detection of calnexin, a membrane protein localized within the endoplasmic reticulum that isn’t present in exosomes, can be obtainable as a unfavorable management for assessing purity of exosome extracts. 

Figure 2. Western detection utilizing ExoBrite™ Western Antibodies and MCF-7 cell and exosome lysates, exhibiting tetraspanin protein enrichment and calnexin depletion in exosomes. Blots had been stained with ExoBrite™ 770/800 Calnexin Antibody (exosome unfavorable management) plus (A) ExoBrite™ 680/700 CD9 Antibody, (B) ExoBrite™ 680/700 CD63 Antibody, or (C) ExoBrite™ 680/700 CD81 Antibody. Lane M: Protein marker. Lane 1: 10 ug cell lysate. Lane 2: 1 ug cell lysate. Lane 3: 1 ug exosome lysate. The blots had been imaged on a LI-COR Odyssey® infrared imaging system.

The launch of ExoBrite™ Antibody Conjugates add to Biotium’s rising line of optimized options for exosome and EV analysis. Biotium additionally affords ExoBrite™ EV Membrane Staining Kits that are designed for vibrant and particular staining of remoted EVs with little to no background. ExoBrite™ EV Membrane Staining Kits can be found with 4 coloration choices for the Pacific Blue™, FITC, PE, and APC channels. Learn extra about Biotium’s novel options for EV analysis.

About Biotium

Biotium is a number one life science reagent producer and provider dedicated to offering high-quality and revolutionary fluorescent instruments that gas scientific discovery. Our collaborative staff of skilled chemists and biologists, who’re on the forefront of fluorescent dye design, apply chemistry-based rules towards producing options for unmet challenges in life science and medical analysis. Since its founding in 2001, Biotium has developed over 30 patented applied sciences which were licensed out to main life science know-how firms worldwide. For extra data, please go to www.biotium.com.


 

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