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Alexa fluor 647 coupled donkey anti mouse secondary antibody

Manufactured by Thermo Fisher Scientific
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Sourced in Germany

Alexa Fluor 647-coupled donkey anti-mouse secondary antibody is a fluorescent-labeled secondary antibody used for detection and visualization of target proteins in various immunoassays and imaging applications. It binds to the primary mouse antibody, allowing for signal amplification and detection.

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Lab products found in correlation

Accuri c6, by BD (3 mentions)

Close spelling variants of this product

Alexa Fluor 647 anti-mouse secondary antibody Alexa Fluor 647 donkey anti-mouse Alexa Fluor 647 anti-mouse antibody Alexa-Fluor-coupled secondary antibodies Alexa Fluor 647 secondary antibody Donkey anti-mouse Alexa-647 Anti-mouse Alexa Fluor 647 Alexa Fluor donkey anti-mouse Alexa Fluor anti-mouse secondary Alexa-coupled secondary antibodies

3 protocols using alexa fluor 647 coupled donkey anti mouse secondary antibody

1

ABCB5+ Cell Isolation Validation

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Following the ABCB5+ cell isolation, but before the enzymatic detachment of the microbeads (which leads to a transient loss of ABCB5 from the cell surface), ABCB5+ cell content was determined after incubation with an Alexa Fluor® 647-coupled donkey anti-mouse secondary antibody (Thermo Fisher, Langenselbold, Germany) targeting the anti-ABCB5 antibody used for the cell isolation. To discriminate between ABCB5+ cells and free bead-antibody complexes, calcein acetoxymethylester (staining metabolically active cells) was added to the cell-secondary antibody suspension before incubation. Fluorescence was measured by flow cytometry (BD Accuri™ C6, Becton Dickinson). Free, unbound bead-antibody complexes were excluded from the ABCB5+ cell content calculation by gating only events with high calcein fluorescence.
Ballikaya S., Sadeghi S., Niebergall-Roth E., Nimtz L., Frindert J., Norrick A., Stemler N., Bauer N., Rosche Y., Kratzenberg V., Pieper J., Ficek T., Frank M.H., Ganss C., Esterlechner J, & Kluth M.A. (2020). Process data of allogeneic ex vivo-expanded ABCB5+ mesenchymal stromal cells for human use: off-the-shelf GMP-manufactured donor-independent ATMP. Stem Cell Research & Therapy, 11, 482.
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2

Quantifying ABCB5+ Cell Content

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After isolation, but before enzymatic detachment of the microbe-ads (which leads to loss of ABCB5 from the cell surface), the ABCB5+ cell content was determined after incubation with an Alexa Fluor 647-coupled donkey anti-mouse secondary antibody (Thermo Fisher Scientific) targeting the anti-ABCB5 antibody used for cell isolation. To allow for discrimination between ABCB5+ cells and free bead-antibody complexes, calcein acetoxymethyl ester was added to the cell suspension before incubation. Fluorescence was measured flow cytometrically (BD Accuri C6). By gating only events with high calcein fluorescence (indicative of viable cells), unbound bead-antibody complexes were excluded from the ABCB5+ cell calculation.
Kerstan A., Niebergall-Roth E., Esterlechner J., Schröder H.M., Gasser M., Waaga-Gasser A.M., Goebeler M., Rak K., Schrüfer P., Endres S., Hagenbusch P., Kraft K., Dieter K., Ballikaya S., Stemler N., Sadeghi S., Tappenbeck N., Murphy G.F., Orgill D.P., Frank N.Y., Ganss C., Scharffetter-Kochanek K., Frank M.H, & Kluth M.A. (2020). Ex vivo-expanded highly pure ABCB5+ mesenchymal stromal cells as Good Manufacturing Practice-compliant autologous advanced therapy medicinal product for clinical use: process validation and first in-human data. Cytotherapy, 23(2), 165-175.
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3

Quantifying ABCB5+ Cell Content

Check if the same lab product or an alternative is used in the 5 most similar protocols
After isolation, but before enzymatic detachment of the microbe-ads (which leads to loss of ABCB5 from the cell surface), the ABCB5+ cell content was determined after incubation with an Alexa Fluor 647-coupled donkey anti-mouse secondary antibody (Thermo Fisher Scientific) targeting the anti-ABCB5 antibody used for cell isolation. To allow for discrimination between ABCB5+ cells and free bead-antibody complexes, calcein acetoxymethyl ester was added to the cell suspension before incubation. Fluorescence was measured flow cytometrically (BD Accuri C6). By gating only events with high calcein fluorescence (indicative of viable cells), unbound bead-antibody complexes were excluded from the ABCB5+ cell calculation.
Kerstan A., Niebergall-Roth E., Esterlechner J., Schröder H.M., Gasser M., Waaga-Gasser A.M., Goebeler M., Rak K., Schrüfer P., Endres S., Hagenbusch P., Kraft K., Dieter K., Ballikaya S., Stemler N., Sadeghi S., Tappenbeck N., Murphy G.F., Orgill D.P., Frank N.Y., Ganss C., Scharffetter-Kochanek K., Frank M.H, & Kluth M.A. (2020). Ex vivo-expanded highly pure ABCB5+ mesenchymal stromal cells as Good Manufacturing Practice-compliant autologous advanced therapy medicinal product for clinical use: process validation and first in-human data. Cytotherapy, 23(2), 165-175.
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