Western Blot for P65 (NF-kB) Expression in Glioma Stem Cell Whole Cell Lysate

February 13, 2020

Centre for Cellular and Molecular Medicine
Ludwig Cancer Research
Postdoctoral Researcher

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Company:

Cell Signaling Technology

Product Name:

NF-κB p65 (D14E12) XP® Rabbit mAb

Catalog Number:

8242

Clone Number:

D14E12

We examined the expression of P65 in glioma cell lines in our study. 1.5 million cells were harvested and lysed in RIPA buffer (150uL) (added 10X protease inhibitor cocktail). Kept on ice for 5 minutes and centrifuged at 13000rpm for 10 min. Supernatant was collected and loaded 60uL of lysate on 10% SDS-PAGE.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Glioma stem cell whole cell lysates

Primary Incubation

1:1000 in 5% BSA, overnight in cold room

Blocking Agent

5%Milk

Secondary Incubation

1:10000 in 5%milk, room temperature for 1 hour

Tertiary Incubation

N/A

Detection

Chemi doc, femto reagent

Results Summary

Observed a single specific band corresponding to P65 protein molecular weight at approximately 65KD size by western blot.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Clean blot with specific band without any non-specific bands

The Bad

N/A

The Bottom Line

Excellent antibody for western blot analysis

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