At 14 DIV, cell cultures were fixed for 15 minutes with 4% paraformaldehyde (wt/vol) in PBS, permeabilized for 10 minutes with 0.2% saponin (wt/vol), and blocked for 60 minutes with 10% goat serum (vol/vol, Abcam) plus 0.1% Tween-20 (vol/vol). Coverslips were then incubated with primary antibodies (anti-GFP: Abcam, 1:1000, chicken; anti-MAP2: EMD, 1:1000, mouse) at room temperature for one hour. Samples were washed three times with 0.02% saponin (wt/vol) in PBS and labeled with Alexa Fluor 488-tagged anti-chicken and Alexa Fluor 546-tagged anti-mouse IgG for 1 hour at room temperature (Invitrogen, 1:400 dilution). Samples were again washed three times and mounted in Fluoromount G mounting medium (SouthernBiotech). Images were obtained using an FV1000 laser-scanning confocal microscope (Olympus) with FV10-ASW 3.1 acquisition software, using a 20x/1.0 NA water objective, under identical laser and gain settings. Images were analyzed using ImageJ (NIH).
Alexa fluor 488 tagged anti chicken
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488-tagged anti-chicken is a fluorescently-labeled secondary antibody used for detection and visualization in immunoassays. The Alexa Fluor 488 dye provides bright, photostable fluorescence for sensitive detection.
Automatically generated - may contain errors
Lab products found in correlation
Fluoromount g mounting medium, by Southern Biotech (2 mentions)
Anti gfp, by Abcam (2 mentions)
Goat serum, by Abcam (2 mentions)
Fv1000 confocal laser scanning microscope, by Olympus (2 mentions)
Alexa fluor 546 tagged anti mouse igg, by Thermo Fisher Scientific (2 mentions)
Fv10 asw 3.1 acquisition software, by Olympus (2 mentions)
2 protocols using alexa fluor 488 tagged anti chicken
1
Immunofluorescence Labeling of Neurons
2
Immunofluorescence Labeling of Neurons
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At 14 DIV, cell cultures were fixed for 15 minutes with 4% paraformaldehyde (wt/vol) in PBS, permeabilized for 10 minutes with 0.2% saponin (wt/vol), and blocked for 60 minutes with 10% goat serum (vol/vol, Abcam) plus 0.1% Tween-20 (vol/vol). Coverslips were then incubated with primary antibodies (anti-GFP: Abcam, 1:1000, chicken; anti-MAP2: EMD, 1:1000, mouse) at room temperature for one hour. Samples were washed three times with 0.02% saponin (wt/vol) in PBS and labeled with Alexa Fluor 488-tagged anti-chicken and Alexa Fluor 546-tagged anti-mouse IgG for 1 hour at room temperature (Invitrogen, 1:400 dilution). Samples were again washed three times and mounted in Fluoromount G mounting medium (SouthernBiotech). Images were obtained using an FV1000 laser-scanning confocal microscope (Olympus) with FV10-ASW 3.1 acquisition software, using a 20x/1.0 NA water objective, under identical laser and gain settings. Images were analyzed using ImageJ (NIH).
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