Cells were cultured on glass coverslips in a 24-well plate and treated with IFN-γ (20 ng/mL), TGF-β1 (20 ng/mL), LPS (1 µg/mL) or CHO F3 clone supernatants (2% and 20%) for 24h. Cells were fixed and permeabilized in 99% ethanol at room temperature for 20 min, then incubated overnight with primary anti-mouse C3a antibodies (dilution 1/500) (BD Biosciences, San Jose, CA, USA, Clone: 187-1162). Cells were subsequently incubated with Alexa Fluor®488-conjugated Goat Anti-Rat secondary antibody (Invitrogen, Thermo Fischer Scientific, Waltham, MA, USA). The nuclear was counterstained with the nuclear fluorochrome 4’,6-diamidino-2-phenylindole (DAPI). Finally, the coverslips were mounted with Vectashield (Vector Labs, Newark, CA, USA). Images were acquired using a ×60 objective on a Nikon Eclipse E2000-U microscope and a Hamamatsu ORCA-ER camera (NIS-Element BR 5.20.00 imaging software, Nikon, Tokyo, Japan).
Alexa fluor 488 conjugated goat anti rat secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Alexa Fluor®488-conjugated Goat Anti-Rat secondary antibody is a fluorescent-labeled antibody that specifically binds to rat primary antibodies. The Alexa Fluor®488 dye provides a green fluorescent signal that can be detected using standard fluorescence detection methods.
Automatically generated - may contain errors
2 protocols using alexa fluor 488 conjugated goat anti rat secondary antibody
1
Immunofluorescence Visualization of C3a
2
Immunostaining of Microtubule Cytoskeleton
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Untreated cells or cells treated with 30 µM nocodazole for 45 or 90 min were fixed with ice-cold methanol for 20 min at −20°C, washed three times with PBS, and incubated in blocking solution (2% BSA in PBS) for 1 h at RT. Coverslips were incubated with anti–α-tubulin antibody (1:500 [MCA77G]; Bio-Rad) in blocking solution for 1 h, washed with PBS, and incubated with Alexa Fluor 488–conjugated goat anti–rat secondary antibody (1:500; Invitrogen) in blocking solution for 1 h at RT. Samples were washed with PBS and mounted on glass slides using ProLong Gold antifade reagent (Invitrogen).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!