Adult midguts were dissected in PBS and fixed for 15 min in PBS containing 4% paraformaldehyde. After fixation, the samples were washed with PBS containing 0.2% Triton X-100 (PBST)and blocked with 1% BSA in PBST. After incubation with primary antibodies anti-β-Gal (1:500, Promega, Z3781) overnight at 4°C. Midguts were washed and then incubated with Alexa fluorescence secondary antibody (1:1000, Thermo Fisher, A32742) and DAPI (1:1000, ThermoFisher, D1306) for 1 h at room temperature, washed, and mounted in Vectashield (Vector, H-1000). Adult midguts, thorax muscles, as well as abdomens containing fat bodies, were dissected in PBS and fixed for 15 min in PBS containing 4% paraformaldehyde. After fixation, the samples were washed with PBST, incubated with Bodipy 493/503 (1 μg/mL, Thermo Fisher, D3922), Phalloidin (1:1000, Thermo Fisher, A12381), or DAPI (1:1000) for 1 h at room temperature, washed, and mounted in Vectashield (Vector, H-1000). Images of fly appearances were performed on a Nikon SMZ18 or Nikon Eclipse Ts2 and confocal images were obtained using a Zeiss LSM880.
Alexa fluorescence secondary antibody
Manufactured by Thermo Fisher Scientific
The Alexa fluorescence secondary antibody is a laboratory tool used to detect and visualize target proteins in biological samples. It is a conjugated antibody that binds to a primary antibody, allowing for the fluorescent labeling and identification of the target protein. The core function of this product is to provide a reliable and sensitive method for protein detection and analysis in various research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti β gal, by Promega (2 mentions)
Eclipse ts2, by Zeiss (2 mentions)
Lsm 880, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (2 mentions)
Vectashield, by Vector Laboratories (2 mentions)
Smz18, by Nikon (2 mentions)
Modular laser system 2, by PerkinElmer (1 mentions)
Anti gfap antibody, by Abcam (1 mentions)
Axiovert 200m microscope, by Zeiss (1 mentions)
Anti lc3 antibody, by Cell Signaling Technology (1 mentions)
Anti map2 antibody, by Merck Group (1 mentions)
Anti nrf2 antibody, by Cell Signaling Technology (1 mentions)
3 protocols using alexa fluorescence secondary antibody
1
Immunohistochemical Analysis of Drosophila Midgut
2
Immunohistochemical Analysis of Drosophila Midgut
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Adult midguts were dissected in PBS and fixed for 15 min in PBS containing 4% paraformaldehyde. After fixation, the samples were washed with PBS containing 0.2% Triton X-100 (PBST)and blocked with 1% BSA in PBST. After incubation with primary antibodies anti-β-Gal (1:500, Promega, Z3781) overnight at 4°C. Midguts were washed and then incubated with Alexa fluorescence secondary antibody (1:1000, Thermo Fisher, A32742) and DAPI (1:1000, ThermoFisher, D1306) for 1 h at room temperature, washed, and mounted in Vectashield (Vector, H-1000). Adult midguts, thorax muscles, as well as abdomens containing fat bodies, were dissected in PBS and fixed for 15 min in PBS containing 4% paraformaldehyde. After fixation, the samples were washed with PBST, incubated with Bodipy 493/503 (1 μg/mL, Thermo Fisher, D3922), Phalloidin (1:1000, Thermo Fisher, A12381), or DAPI (1:1000) for 1 h at room temperature, washed, and mounted in Vectashield (Vector, H-1000). Images of fly appearances were performed on a Nikon SMZ18 or Nikon Eclipse Ts2 and confocal images were obtained using a Zeiss LSM880.
3
Immunofluorescence Staining of Cells
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The cells were seeded on 12-well plates or coverslips coated with poly-D-lysine hydrobromide, fixed in 4% paraformaldehyde, and washed with 0.1% Triton X-100 in PBS. The fixed cells were blocked with 10% normal goat serum for 1 h at room temperature followed by incubation with primary antibodies at 4°C overnight. Cells were incubated with Alexa fluorescence secondary antibody (Thermo Scientific) for 1 h in the dark. Cells were then washed and kept in the PBS. Coverslips were further mounted with the antifading mounting reagent (Thermo Scientific). Images were taken using the Modular Laser System 2.0 (PerkinElmer) with the Nikon ECLIPSE TE2000-E confocal system (Nikon) or the Zeiss Axiovert 200 M microscope (Zeiss). Antibodies used in this research included the following: anti-GFAP antibody (number: ab2760, 1 : 5000; number: MAB360, 1 : 400) (Abcam; Millipore), anti-MAP2 antibody (number: MAB3418, 1 : 200) (Millipore), anti-LC3 antibody (number: 12741, 1 : 100) (Cell signaling), anti-p62 antibody (number: PM045, 1 : 500) (MBL), and anti-Nrf2 antibody (number: 12721, 1 : 100) (Cell signaling).
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