Western Blot of TAP-SPT16 in S. cerevisiae Whole-Cell Lysates

February 18, 2019

Laboratory of t-RNA Transcription
Centre for Cellular and Molecular Biology
Research Associate

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen

Product Name:

TAP Tag Polyclonal Antibody

Catalog Number:

CAB1001

SPT16 is a component of FACT complex (SPT16-Pob3) in Saccharomyces cerevisiae. FACT complex is involved in the regulation of chromatin-associated processes such as DNA replication and repair, etc. Our aim was to examine the expression of TAP-tagged SPT16 protein in whole-cell lysates of TAP-SPT16 expressing S. cerevisiae cells. We isolated protein from cells by bead beating and loaded on SDS-PAGE and performed western blots using this antibody.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Saccharomyces cerevisiae cell lysate

Primary Incubation

1:4000, 2 hour at room temperature

Blocking Agent

5% BSA

Secondary Incubation

1:10000, anti-rabbit HRP conjugated antibody, 1 hr at room temperature

Tertiary Incubation

N/A

Detection

ECL reagent, Chemi doc

Results Summary

We probed the blot with TAP antibody and observed a band corresponding to TAP-SPT16 at 140 kDa in cell lysates suggesting the expression of TAP-SPT16 in cells expressing TAP-SPT16 gene.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Good antibody for performing western blots

The Bad

A few non-specific bands after long exposure

The Bottom Line

Overall an excellent antibody to perform western blots and easy to optimize.

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