Immunoblotting Analysis of Extracellular Vesicle Markers

Immunoblotting was performed as previously described [6 (link)]. Proteins were detected using primary anti-CD9 (ab92726, Abcam, Cambridge, MA), anti-TSG101 (T5701, MilliporeSigma, St. Louis, MO), anti-Alix (ab186429, Abcam), anti-iNOS (13120S, Cell Signaling), anti-Arg-1 (93668S, Cell Signaling), anti-GAPDH (437,000, Thermo Fisher Scientific), anti-P (phospho)-Akt Ser473 (4060, Cell Signaling), anti-P-Akt Thr308 (13,038, Cell Signaling), and anti-Akt (4685, Cell Signaling) antibodies. Exposure of the resultant protein bands was performed with an ImageQuant LAS 4000 Luminescent Image Analyzer (GE Healthcare; Chicago, IL).

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Zhu D., Johnson T.K., Wang Y., Thomas M., Huynh K., Yang Q., Bond V.C., Chen Y.E, & Liu D. (2020). Macrophage M2 polarization induced by exosomes from adipose-derived stem cells contributes to the exosomal proangiogenic effect on mouse ischemic hindlimb. Stem Cell Research & Therapy, 11, 162.

Publication 2020

ab92726 ab186429, anti-Akt (4685, ImageQuant LAS 4000 Luminescent Image Analyzer

Antibodies Gapdh Inos Luminescent Protein Tsg101

Corresponding Organization : Morehouse School of Medicine

Other organizations : National Eye Institute, Louisiana State University, Louisiana State University Health Sciences Center New Orleans, University of Michigan–Ann Arbor

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Variable analysis

independent variables

Primary antibodies used for protein detection: anti-CD9, anti-TSG101, anti-Alix, anti-iNOS, anti-Arg-1, anti-GAPDH, anti-P (phospho)-Akt Ser473, anti-P-Akt Thr308, and anti-Akt

dependent variables

Protein expression levels detected using the primary antibodies

control variables

Exposure of the resultant protein bands was performed with an ImageQuant LAS 4000 Luminescent Image Analyzer (GE Healthcare; Chicago, IL)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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