Not K48 Linkage Specific! Also Recognizes K63 Linkages

University of Kansas Medical Center

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Company:

Cell Signaling Technology

Product Name:

K48-linkage Specific Polyubiquitin Antibody

Catalog Number:

4289

Clone Number:

I do a lot of ubiquitin-proteasome system work in yeast (S. cerevisiae), including looking at (poly)ubiquitination.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Yeast whole cell lysates and polyubiquitin standards

Primary Incubation

1:1000, overnight 4 degrees

Blocking Agent

5% non-fat milk in TBST

Secondary Incubation

HRP conjugated anti-rabbit IgG

Tertiary Incubation

n/a

Detection

Immobilon Forte HRP substrate, chemiluminescence

Results Summary

This antibody is supposed to be a K48-linkage specific ubiquitin antibody, but it recognized a standard 2- to 7-mer of K63-linked polyubiquitin. The image shows a comparison between this antibody, a pan-ubiquitin antibody, and another CST antibody for K48-linked polyUb that works.

Image Gallery

Summary

The Good

nothing

The Bad

It is not specific to K48 linkages

The Bottom Line

Validate this antibody against K63-linked polyUb standards before use or avoid using it for K48-linked polyUb altogether. There are alternatives (like CST #8081).

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