Reliable and Precise IC50 Values

Cancer System Imaging (Diagnostic Imaging Division)
MD Anderson Cancer Center - University of Texas
Lab Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

R&D Systems

Product Name:

PARP/Apoptosis Universal Colorimetric Assay Kit

Catalog Number:

4677-096-K

Clone Number:

N/A

In our lab we study PARP-1 that is a nuclear protein and mediates Poly-ADP-ribosylation o nuclear proteins (as it is, and histones for example), a process that occurs in response to DNA damage. Inhibiting PARP has shown to promote chemo/radiosensitization of tumors in the conventional therapy of malignant tumors. Furthermore, PARP1 inhibitors can be used as independent, effective drugs against tumors with broken DNA repair mechanisms.Our aim was doing a vitro screening of a candidate PARP-1 inhibitor in order to determine its IC50 value. As we know PARP-1 release a biotinylated poly (ADP-ribose) that attach the 96-wells surface coated with histones. Detections of PARP-1 was made through the absorbance (at 450nm) of the colorimetric HRP substrate of Strep-HRP (a biotin binding protein). We found a reliable value of IC50. We will proceed testing other inhibitors with this kit!

Experimental Design and Results Summary

Applications

ELISA

Sample

Inhibitor of PARP-1

Primary Incubation

N/A

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Absorbance at 450nm

Results Summary

In our lab we study PARP-1 that is a nuclear protein and mediates Poly-ADP-ribosylation o nuclear proteins (as it is, and histones for example), a process that occurs in response to DNA damage. Our aim was doing a vitro screening of a candidate PARP-1 inhibitor in order to determine its IC50 value. Inhibiting PARP has shown to promote chemo/radiosensitization of tumors but can be used as independent, effective drugs against tumors with broken DNA repair mechanisms.As we know PARP-1 release a biotinylated poly (ADP-ribose) that attach the 96-wells surface coated with histones. Detections of PARP-1 was made through the absorbance (at 450nm) of the colorimetric HRP substrate of Strep-HRP (a biotin binding protein). We found a reliable value of IC50. We will proceed testing other inhibitors with this kit!

DOI or PMID #

26483957

Additional Notes

Photo attached (Histone-Coated Clear Strip Wells, 96-well _ Cat. No. 4677-096-P)

Image Gallery

Summary

The Good

Reliable and precise results (IC50 values), easy to use.

The Bad

Confusing protocol (units of measure used to indicate the same substance change).

The Bottom Line

Clear and reliable results, easy to follow protocol.

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