Cells were washed, blocked with 2% BSA and incubated with 1:1000 of VesB antiserum that was pre-incubated with ΔvesABC cells to remove cross-reactive antibodies. Following incubation with 1:1000 of Alexa Fluor 488 F(ab′)2 goat anti-rabbit immunoglobulin G (Invitrogen) and washing, fluorescence was measured (Ex 495 nm/ Em 519 nm). The cells were also visualized by differential interference contrast and fluorescent microscopy using a Nikon Eclipse 90i fluorescence microscope equipped with a Nikon Plan Apo VC 100× (1.4 numerical aperture) oil immersion objective and a CoolSNAPHQ2 digital camera as previously described [57 (link)].
Alexa fluor 488 f ab 2 goat anti rabbit immunoglobulin g
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488 F(ab′)2 goat anti-rabbit immunoglobulin G is a secondary antibody conjugate used for the detection and visualization of rabbit primary antibodies in various immunoassay and microscopy applications. The Alexa Fluor 488 fluorescent dye is attached to the F(ab′)2 fragment of the goat anti-rabbit IgG antibody, providing a bright and stable fluorescent signal.
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2 protocols using alexa fluor 488 f ab 2 goat anti rabbit immunoglobulin g
1
Visualization of VesB Localization
2
Detecting Bacterial Virulence Factors
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Cells were washed, blocked with 2% BSA, and incubated with 1:1,000 of VesB antiserum that was pre-incubated with ΔvesABC cells to remove cross-reactive antibodies. Following incubation with 1:1,000 of Alexa Fluor 488 F(ab′)2 goat anti-rabbit immunoglobulin G (Invitrogen) and washing, fluorescence was measured (Ex 488 nm/Em 525 nm). The results were normalized to the fluorescence intensity of the same strain carrying an empty vector as described (10) (link).
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