DENV-2 Immunostaining with Anti -dengue Type 2

Biomedical Sciences
Texas Tech University Health Science Center, El paso Texas
Postdoctoral Research Fellow

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Company:

US Biological

Product Name:

Anti -dengue Type 2

Catalog Number:

D2800-10

Dengue virus (DENV-2) infected HeLa cells were immunostained for P- bodies and DENV-2. Cells grown on cover slips were washed in PBS, fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and blocked (2% bovine serum albumin, 5% normal horse serum, and 10 mM glycine in phosphatebuffered saline). The cells were then incubated with a rabbit anti -DCP1a antibody (Abcam) and DENV-2 specific antibodies (US Biologicals), followed by incubation with appropriate secondary antibodies (AlexaFluor 594-conjugated anti-rabbit antibody and fluorescein isothiocyanate-conjugated anti-mouse antibody (Invitrogen). The cells were visualized for DENV-2 and DCP1a staining using a Nikon Eclipse fluorescence microscope. The findings were published in PLOS One Journal (PLoS One. 2011;6(11):e27551).

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

DENV-2 infected HeLa cells

Primary Incubation

1:500, 1hr at RT or O/N at 4 degrees Celsius in PBS

Blocking Agent

2% bovine serum albumin, 5% normal horse serum, and 10 mM glycine in phosphate buffered saline

Secondary Incubation

1:1000 fluorescein isothiocyanate-conjugated anti-mouse antibody for 1 hour at RT in PBS

Tertiary Incubation

NA

Detection

Fluorescein isothiocyanate-conjugated anti-mouse antibody, visualized using a Nikon Eclipse fluorescence microscope

Results Summary

The anti DENV-2 antibody gave specific signal in DENV-2 infected HeLa cells and no signal was observed in uninfected cells. The findings were published in PLOS One Journal (PLoS One. 2011;6(11):e27551).

Additional Notes

None

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Summary

The Good

DENV-2 specific reactivity

The Bad

None

The Bottom Line

This is a specific monoclonal antibody that worked really well in immunofluorescence. However, we did not test it in other applications so I cannot comment on that part.

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