FZD7 Abs to verify the transfection of HEK 293 cells

Biology
University of Ottawa
PhD student

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Company:

Novus Biologicals

Product Name:

Frizzled-7 Antibody (151143) [Allophycocyanin]

Catalog Number:

FAB1981A

Info: View Product View Product Specs

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Our laboratory selects aptamers for various cell receptors. To test the specificity of aptamer binding to white blood cells or transfected HEK-293 or MDA-MB-231, we use receptor specific antibodies. Aptamers can be labeled with fluorescent dyes or biotinylated. To detect biotinylated aptamers conjugated streptavidin is used. All cells are stained with propidium iodide or DRAQ7 to distinguish live from dead cells. HEK 293 cells were transfected with FZD7 and later used for aptamer selection. Abs were used to verify the transfection of HEK 293 cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

300000 cells in 100 ul

Primary Incubation

90 minutes, 5 ul per sample (100 ul), room temperature, DPBS with Ca and MG

Blocking Agent

1% FBS in DPBS with Ca and Mg

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Beckman Gallios flow cytometer

Results Summary

Transfected cells gave statistically higher signal than non-transfected cells.

Additional Notes

None

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Summary

The Good

Abs gave good signal

The Bad

Blocking needs to be adjusted to prevent non-specific binding

The Bottom Line

In our experiment Abs worked well to verify the transfection.

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