Alexa fluor 450 streptavidin

Manufactured by Thermo Fisher Scientific

Alexa Fluor 450 streptavidin is a fluorescent labeling reagent used in various bioassays and imaging applications. Streptavidin, a protein derived from the bacterium Streptomyces, has a high affinity for the small molecule biotin. The Alexa Fluor 450 dye is covalently attached to the streptavidin, providing a fluorescent label that can be used to detect and visualize biotinylated biomolecules.

Automatically generated - may contain errors

Lab products found in correlation

Anti mouse igg2b antibody staining, by Thermo Fisher Scientific (2 mentions) Soluble biotinylated anti igm, by Southern Biotech (2 mentions) Ebioy ae, by Thermo Fisher Scientific (2 mentions)

2 protocols using alexa fluor 450 streptavidin

B Cell Antigen Internalization and Presentation

Check if the same lab product or an alternative is used in the 5 most similar protocols

For internalization assays, purified B cells were loaded with IgM coated beads or with soluble biotinylated anti-IgM (Southern Biotech) on ice for 30 min. Cells were then washed with PBS 2% FCS to remove excess antigen, and then incubated for 5–30 min at 37°C. After fixation with 2% paraformaldehyde, noninternalized beads were detected with Alexa Fluor 450 streptavidin (eBioscience).
To detect antigen presentation, B cells loaded with Ea peptide and IgM-coated beads were incubated between 3 and 5 h at 37°C, and then fixed in 2% formaldehyde. These cells were then stained with anti-MHCII/Ea antibody (eBioY-Ae; eBioscience), followed by anti–mouse IgG2b antibody staining (Life Technologies) for detection.

Burbage M., Keppler S.J., Gasparrini F., Martínez-Martín N., Gaya M., Feest C., Domart M.C., Brakebusch C., Collinson L., Bruckbauer A, & Batista F.D. (2015). Cdc42 is a key regulator of B cell differentiation and is required for antiviral humoral immunity. The Journal of Experimental Medicine, 212(1), 53-72.

+ Open protocol

+ Expand

Internalization and Antigen Presentation Assay for B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For internalization assays, purified B cells were loaded with IgM coated fluorescently-labelled microspheres or with soluble biotinylated anti-IgM (Southern Biotech) on ice for 30 min. Cells were then washed with PBS 2% FCS to remove excess antigen and then incubated for 5 to 30 min at 37°C. After fixation with 2% paraformaldehyde, non-internalised beads or anti-IgM were detected with AlexaFluor450 streptavidin (eBioscience, San Diego, California). In case of bead internalisation, bead-positive cells were selected on the basis of the bead fluorescence.
To detect antigen presentation, B cells loaded with Eα peptide and IgM coated microspheres were incubated between 3 and 5 hr at 37°C and then fixed in 2% formaldehyde. These cells were then stained with anti-MHCII/Eα antibody (eBioY-Ae, eBioscience), followed by anti-mouse IgG2b antibody staining (Life Technologies) for detection.

Burbage M., Gasparrini F., Aggarwal S., Gaya M., Arnold J., Nair U., Way M., Bruckbauer A, & Batista F.D. (2018). Tuning of in vivo cognate B-T cell interactions by Intersectin 2 is required for effective anti-viral B cell immunity. eLife, 7, e26556.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!