Phospho-Rb Antibody

Pathology
University of Kansas Medical Center
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-Rb (Ser780) (D59B7)

Catalog Number:

8180

We are interested in mechanisms of resistance to targeted therapies in ovarian cancer. This experiment was designed to determine differential Rb1 protein expression and post-translational modifications in ovarian cancer cells lines resistant to a therapy, compared with therapy-sensitive controls.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Human ovarian cancer cell line lysates

Primary Incubation

1:1000 dilution in 5% milk in TBS-T, overnight at 4 degrees C

Blocking Agent

5% milk in TBS-T

Secondary Incubation

1:10,000 in 5% milk in TBS-T, 30 minutes at RT

Tertiary Incubation

N/A

Detection

Chemiluminescence substrate for HRP-labeled secondary antibodies

Results Summary

We observed multiple bands on the blot, which may be due to non-specificity of the antibody or modified protein (identity of bands remains unclear). The staining was very strong, suggesting the antibody can be further diluted to help more clearly reflect differences (if any) in band intensity.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Very strong bands observed

The Bad

Multiple nonspecific bands observed

The Bottom Line

Great antibody that can be used at dilute concentrations. However, it is unclear if the observed bands are biologically relevant or an artifact.

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