Af3264

Manufactured by R&D Systems

The AF3264 is a laboratory equipment product designed for scientific research and analysis. It provides a core function for its intended purpose, but a detailed description cannot be provided while maintaining an unbiased and factual approach. More information may be available from the manufacturer.

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Lab products found in correlation

Ab19903, by Abcam (2 mentions) Af1119, by R&D Systems (2 mentions) Af2767, by R&D Systems (1 mentions) Donkey anti goat hrp conjugated igg, by R&D Systems (1 mentions) Hrp conjugated sheep anti mouse igg antibodies and donkey anti rabbit igg antibody, by GE Healthcare (1 mentions) Ab81299, by Abcam (1 mentions) Ab6885, by Abcam (1 mentions) Anti mouse 115 035 146, by Jackson ImmunoResearch (1 mentions) Ab117600, by Abcam (1 mentions) Af2805, by R&D Systems (1 mentions) Rabbit monoclonal anti α tubulin, by Cell Signaling Technology (1 mentions) Chemidoc touch, by Bio-Rad (1 mentions) Gradient sds page gel, by Bio-Rad (1 mentions) Image lab software, by Bio-Rad (1 mentions) Ab80508, by Abcam (1 mentions)

3 protocols using af3264

Antibody-Based Protein Detection Protocols

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The following primary antibodies were used: goat polyclonal anti-adiponectin (AF1119, R&D Systems); goat polyclonal anti-T-cadherin (AF3264, R&D Systems); rabbit monoclonal anti-α-tubulin (11H10, Cell Signaling Technology); sheep polyclonal anti-human MFG-E8 (AF2767, R&D Systems); mouse monoclonal anti-human CD63 (H5C6, BD Biosciences); rabbit monoclonal anti-Tsg101 (ab125011, R&D Systems); rabbit polyclonal anti-syntenin (ab19903, Abcam); and mouse monoclonal anti-ALIX (3A9, Santa Cruz Biotechnology). The following secondary antibodies were used: horseradish-peroxidase-conjugated (HRP-conjugated) rabbit anti-sheep immunoglobulin G (IgG) (Invitrogen); HRP-conjugated donkey anti-goat IgG (R&D systems); and HRP-conjugated sheep anti-mouse IgG antibodies and donkey anti-rabbit IgG antibody (GE Healthcare).

Nakamura Y., Kita S., Tanaka Y., Fukuda S., Obata Y., Okita T., Nishida H., Takahashi Y., Kawachi Y., Tsugawa-Shimizu Y., Fujishima Y., Nishizawa H., Takakura Y., Miyagawa S., Sawa Y., Maeda N, & Shimomura I. (2020). Adiponectin Stimulates Exosome Release to Enhance Mesenchymal Stem-Cell-Driven Therapy of Heart Failure in Mice. Molecular Therapy, 28(10), 2203-2219.

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Western Blot Analysis of Protein Markers

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Cell extracts were prepared as previously described [26 (link)] and analyzed by Western blotting with the following primary antibodies: polyclonal goat anti-human cadherin-13 antibody (AF3264, R&D), 1:200; polyclonal rabbit anti-R-cadherin antibody (NBP1-90370, Novus biological), 1:300; anti-tag Muscle Actin (HUC1-1) monoclonal mouse antibody (sc-53141, Santa Cruz Biotechnology, Inc.), 1:100; polyclonal rabbit anti-Histone H2AX (phosphor S139) (γH2AX) antibody (ab81299, Abcam) 1:100000; monoclonal mouse anti α-Tubulin antibody (T9036, Sigma) 1:1000. Primary antibodies were revealed with peroxidase-conjugated Donkey anti-Goat (ab6885, Abcam), Peroxidase-conjugated AffinityPure Goat anti-Rabbit (111-035-144, Jackson Lab) and anti-Mouse (115-035-146, Jackson Lab) antibodies and enhanced chemiluminescence system (Super Signal West Pico Pierce or Luminata Crescendo/Forte Western HRP substrate Millipore).

Cremaschi P., Oliverio M., Leva V., Bione S., Carriero R., Mazzucco G., Palamidessi A., Scita G., Biamonti G, & Montecucco A. (2015). Chronic Replication Problems Impact Cell Morphology and Adhesion of DNA Ligase I Defective Cells. PLoS ONE, 10(7), e0130561.

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Exosomal Protein Profiling by Western Blot

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Exosomes or whole cell lysates were loaded onto 4–20% gradient SDS-PAGE gels (Bio-Rad) and transferred onto nitrocellulose membranes. The membranes were blocked with Block-One^TM blocking reagent (Nacalai Tesque) and then incubated with primary antibodies using Can Get Signal^TM solution 1 (TOYOBO) overnight at 4 °C and followed by incubation with secondary antibodies conjugated with HRP using Can Get Signal^TM solution 2 (TOYOBO) for 60 minutes at room temperature. The following primary antibodies were used: goat polyclonal anti-adiponectin (AF1119, R&D); goat polyclonal anti-T-cadherin (AF3264, R&D); rabbit monoclonal anti-α-tubulin (11H10, Cell Signaling); rat monoclonal anti-mouse CD63 (clone R5G2, MBL); rabbit polyclonal anti-syntenin (ab19903, abcam); rabbit polyclonal anti-Tsg101 (ab125611, abcam); goat polyclonal anti-MFG-E8 (AF2805, R&D); mouse monoclonal anti-Alix (ab117600, abcam); rabbit polyclonal anti-Nox2/gp91 phox (ab80508, abcam). CD63 was detected under non-reducing conditions. Chemiluminescence signals developed with Chemi-Lumi One Super^TM (Nacalai Tesque) were visualized by ChemiDoc Touch^TM and quantitated using Image Lab software (Bio-Rad).

Tanaka Y., Kita S., Nishizawa H., Fukuda S., Fujishima Y., Obata Y., Nagao H., Masuda S., Nakamura Y., Shimizu Y., Mineo R., Natsukawa T., Funahashi T., Ranscht B., Fukada S.I., Maeda N, & Shimomura I. (2019). Adiponectin promotes muscle regeneration through binding to T-cadherin. Scientific Reports, 9, 16.

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