Anti cd81

Manufactured by R&D Systems

Sourced in United States

Anti-CD81 is a laboratory reagent used for the detection and analysis of CD81, a cell surface protein. It can be used in various applications such as flow cytometry and immunoprecipitation to study the expression and function of CD81 in different cell types.

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Lab products found in correlation

Anti cd63, by BD (2 mentions) Anti cd9, by R&D Systems (2 mentions) Sc 9996, by Santa Cruz Biotechnology (1 mentions) Anti flag hrp, by Merck Group (1 mentions) Anti calnexin, by Cell Signaling Technology (1 mentions) Nel104001, by PerkinElmer (1 mentions) Anti hsp90, by Cell Signaling Technology (1 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (1 mentions)

3 protocols using anti cd81

High-Resolution Imaging of Single Extracellular Vesicles

Cited 2 times

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To obtain sub‐diffraction limit resolution images of single Evs, 1 × 10⁹ Evs in 1 μl were stained with a ‘tetramix’ of anti‐CD9, (R&D Systems, USA), anti‐CD63, (Becton Dickinson, USA) and anti‐ CD81 (R&D Systems, USA), all conjugated to AlexaFluor 647 (Gomes et al., 2022 (link)) or with antibodies supplied in the EV profiler Kit (Oxford Nanoimaging, UK) including anti‐CD9‐ATTO488, anti‐CD63‐Cy3 and anti‐CD81‐AlexaFluor647. Samples were processed according to manufacturer's instructions to immobilise the stained Evs on the chips provided with the ONI EV profiler kit. Three fields of view were recorded for each sample using direct stochastic optical reconstruction microscopy (dSTORM) on a Nanoimager S Instrument (Oxford Nanoimaging, UK) using 40%, 20% and 20% power for the 488 nm, 561 nm and 640 nm laser, respectively. For localisation mapping, 2500 images were recorded per channel. To quantify the images and to detect colocalisations, we either used the EVAnalyzer software as described in the manuscript, or the CODI platform (Oxford Nanoimaging, UK) as described previously (Wolf et al., 2022 (link)).

Schürz M., Danmayr J., Jaritsch M., Klinglmayr E., Benirschke H.M., Matea C., Zimmerebner P., Rauter J., Wolf M., Gomes F.G., Kratochvil Z., Heger Z., Miller A., Heuser T., Stanojlovic V., Kiefer J., Plank T., Johnson L., Himly M., Blöchl C., Huber C.G., Hintersteiner M, & Meisner‐Kober N. (2022). EVAnalyzer: High content imaging for rigorous characterisation of single extracellular vesicles using standard laboratory equipment and a new open‐source ImageJ/Fiji plugin. Journal of Extracellular Vesicles, 11(12), 12282.

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Western Blot Analysis of Protein Expression

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Whole cells were trypsinized and harvest subjected to Laemmli sampling buffer and boiled for at 95°C for 10 min. The protein extracts were separated on SDS-PAGE gels, transferred to PVDF membranes and incubated with indicated antibodies. The antibodies were purchased from commercial sources: anti-FLAG HRP (1:1000, Sigma, A8592), anti-HSP90 (1:1000, Cell Signaling Technologies, 4877S), anti-CD63 (1:600, BD Biosciences, 556019), anti-CD81 (1:600, R&D Systems, MAB4615), anti-Calnexin (1:1000, Cell Signaling Technologies, 2679T), anti-β-Tublin (1:1000, Proteintech, 10094-1-AP), anti-Mettl3 (1:1000, Proteintech, 15073-1-AP), anti-GFP (1:500, Santa Cruz Biotechnology, sc-9996), ALTO antiserum [13 (link)] (1:5000), and TSPyV pAbMT (1:100) [14 (link)]. Membranes were then incubated with the appropriate HRP-conjugated secondary antibody (1:000, anti-mouse IgG, Cell Signaling Technologies, 7076P2; 1:1000, anti-rabbit IgG, ThermoFisher, G21234) and developed with an ECL system (Perkin Elmer, NEL104001).

Yang R., Lee E.E., Kim J., Choi J.H., Kolitz E., Chen Y., Crewe C., Salisbury N.J., Scherer P.E., Cockerell C., Smith T.R., Rosen L., Verlinden L., Galloway D.A., Buck C.B., Feltkamp M.C., Sullivan C.S, & Wang R.C. (2021). Characterization of ALTO-encoding circular RNAs expressed by Merkel cell polyomavirus and trichodysplasia spinulosa polyomavirus. PLoS Pathogens, 17(5), e1009582.

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Extracellular Vesicle Immunophenotyping

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EVs were bound to CD63-conjugated magnetic beads (Dynabeads 10606D, Thermo Fisher Scientific) at 4℃ overnight. Beads were washed and incubated with anti-CD9 (R&D Systems), anti-CD81 (R&D Systems), and anti-CD73 (R&D Systems), or their respective isotype controls for 1 h at RT at 700 rpm. After 3 × washes in 0.1% BSA/PBS solution, the beads were resuspended and analyzed using an Attune NxT Flow Cytometer (Thermo Fisher Scientific).

Lyu N., Knight R., Robertson S.Y.T., Dos Santos A., Zhang C., Ma C., Xu J., Zheng J, & Deng S.X. (2022). Stability and Function of Extracellular Vesicles Derived from Immortalized Human Corneal Stromal Stem Cells: A Proof of Concept Study. The AAPS journal, 25(1).

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