Guts were dissected in PBS and transferred into 4% PFA immediately after dissection and staining was performed on an orbital shaker. After 45 min of fixation guts were washed once with PBS for 10 min. Antibodies were diluted in 0.5% PBT + 5% normal goat serum. The incubation with primary antibodies (1:250 anti-Dlg-1 [mouse; Developmental studies Hybridoma Bank (DSHB)]; 1:50 anti-Pros [mouse; DSHB]; 1:2000 anti-pH3 [rabbit; Merck Millipore, 06–570]; 1:50 anti-EcR common Ag10.2 [mouse; DSHB]; 1:500 anti-HA High Affinity 3F10 [rat; Merck, Sigma-Aldrich]; 1:1500 anti-ß-Galactosidase preabsorbed [rabbit; Cappel Research Products]) was performed at 4°C over night. After washing with PBS guts were incubated with secondary antibodies (1:500 Goat anti-MouseAlexa647 [Invitrogen]; 1:500 Goat anti-RatAlexa647 [Invitrogen]; 1:500 Goat anti-RabbitAlexa647 [Invitrogen]) and DAPI (1:1000; 100 µg/ml stock solution in 0.18 M Tris pH 7.4; DAPI No. 18860, Serva, Heidelberg) for at least 3 hr at RT. Guts were washed a last time with PBS prior to mounting in Fluoromount-G Mounting Medium (Electron Microscopy Sciences).
Goat anti ratalexa647
Manufactured by Thermo Fisher Scientific
Goat anti-RatAlexa647 is a secondary antibody conjugated with Alexa Fluor 647 dye. It is used to detect and visualize rat primary antibodies in various immunodetection techniques, such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse alexa 647, by Thermo Fisher Scientific (2 mentions)
Fluoromount g mounting medium, by Electron Microscopy Sciences (2 mentions)
Goat anti rabbitalexa647, by Thermo Fisher Scientific (2 mentions)
Dapi no 18860, by Serva Electrophoresis (2 mentions)
Anti dlg1, by Developmental Studies Hybridoma Bank (1 mentions)
Anti ha high affinity 3f10, by Merck Group (1 mentions)
Anti ph3, by Merck Group (1 mentions)
2 protocols using goat anti ratalexa647
1
Immunostaining of Drosophila Gut
2
Immunofluorescence Staining of Gut Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Guts were dissected in PBS and transferred into 4% PFA immediately after dissection and staining was performed on an orbital shaker. After 45 min of fixation guts were washed once with PBS for 10 min. ) was performed at 4°C over night. After washing with PBS guts were incubated with secondary antibodies (1:500 Goat anti-MouseAlexa647 [Invitrogen]; 1:500 Goat anti-RatAlexa647
[Invitrogen]; 1:500 Goat anti-RabbitAlexa647 [Invitrogen]) and DAPI (1:1000; 100 µg/ml stock solution in 0.18 M Tris pH 7.4; DAPI No. 18860, Serva, Heidelberg) for at least 3 h at RT. Guts were washed a last time with PBS prior to mounting in Fluoromount-G™ Mounting Medium (Electron Microscopy Sciences).
[Invitrogen]; 1:500 Goat anti-RabbitAlexa647 [Invitrogen]) and DAPI (1:1000; 100 µg/ml stock solution in 0.18 M Tris pH 7.4; DAPI No. 18860, Serva, Heidelberg) for at least 3 h at RT. Guts were washed a last time with PBS prior to mounting in Fluoromount-G™ Mounting Medium (Electron Microscopy Sciences).
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