For immunofluorescent staining of focal adhesion structures, 50,000 (for D line) or 100,000 (for B line) cells were plated on cover slips in 6-well plates, and were grown for 2 days before paraformaldehyde fixation. The fixed cells were then stained with mouse anti-paxillin (BD #610052, 1:500) or mouse anti-vinculin (Sigma-Aldrich V9131, 1:800), along with DAPI (Life Technologies) and Alexa Fluor 555 Phalloidin (ThermoFisher Scientific). The secondary antibody used was donkey anti-mouse IgG (H+L) Alexa Fluor 647 conjugate (ThermoFisher Scientific, 1:250). The stained cells were imagined with Applied Precision DeltaVision Spectris Imaging microscope, and images were deconvoluted with the Softworx deconvolution software.
Donkey anti mouse igg h l alexa fluor 647 conjugate
Manufactured by Thermo Fisher Scientific
Donkey anti-mouse IgG (H+L) Alexa Fluor 647 conjugate is a secondary antibody reagent. It is conjugated to the Alexa Fluor 647 dye, which has excitation and emission maxima at 650 nm and 665 nm, respectively. This reagent can be used to detect and visualize mouse immunoglobulins in various applications.
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Lab products found in correlation
2 protocols using donkey anti mouse igg h l alexa fluor 647 conjugate
1
Immunofluorescent Staining of Focal Adhesions
2
Immunostaining Drosophila Wing and Embryo
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Primary antibodies: mouse anti-β-catenin (Clone 14/Beta-Catenin) (1:500, BD Biosciences Cat. # 610154), mouse anti-Wingless (1:25, DSHB Cat. # 4D4 (supernatant) for D. melanogaster wing imaginal discs and 1:200, DSHB Cat. # 4D4 (concentrate) for embryos), guinea pig anti-Senseless (1:1000; Nolo et al., 2000 (link)), mouse anti-Engrailed/Invected (1:100, DSHB Cat. # 4D9 (concentrate)).
Secondary antibodies: donkey anti-mouse IgG (H+L) Alexa Fluor 647 conjugate (1:250, Thermo Fisher Scientific Cat. # A-31571), donkey anti-mouse IgG (H+L) Alexa Fluor 555 conjugate (1:400, Thermo Fisher Scientific Cat. # A-31570), goat anti-guinea pig IgG (H+L) highly cross-adsorbed, Alexa Fluor 488 conjugate (1:400, Thermo Fisher Scientific Cat. # A-11073).
Secondary antibodies: donkey anti-mouse IgG (H+L) Alexa Fluor 647 conjugate (1:250, Thermo Fisher Scientific Cat. # A-31571), donkey anti-mouse IgG (H+L) Alexa Fluor 555 conjugate (1:400, Thermo Fisher Scientific Cat. # A-31570), goat anti-guinea pig IgG (H+L) highly cross-adsorbed, Alexa Fluor 488 conjugate (1:400, Thermo Fisher Scientific Cat. # A-11073).
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