Good Antibody to Detect Human Twist Protein

September 12, 2018

The Cell and Molecular Biology
The Scripps Research Institute
Assistant Professor

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Company:

Santa Cruz Biotechnology

Product Name:

twist (Twist2C1a)

Catalog Number:

sc-81417

Twist (also known as twist-related protein 1, or TWIST1) belongs to the group of basic helix-loop-helix proteins regulating cell differentiation during embryogenesis. Twist also plays a critical role in cancer metastasis and is known to be over-expressed in metastatic carcinomas. We analyzed Twist levels in human carcinoma cell lines in comparison with the human cells over-expressing Twist protein. In western blotting assays, the antibody sc-81417 from Santa Cruz was capable of recognizing the twist-related protein band with an expected mol. wt. of ~25-28 kDa, though the antibody also bound to several non-specific protein bands of larger mol. wt.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Whole cell lysates of human carcinoma cells

Primary Incubation

Antibody at a 1:1,000 dilution, ON incubation with gentle rocking in a cold room

Blocking Agent

5% milk in PBS-0.05% Tween 20

Secondary Incubation

Stabilized goat anti mouse IgG HRP conjugate, 1:5,000 in blocking buffer

Tertiary Incubation

N/A

Detection

West Pico

Results Summary

In our hands, this mouse monoclonal antibody recognized the protein band with an expected mol. wt. of twist (~25-28 kDa). However, the antibody also produced numerous non-specific bands, which is unfortunately common for transcription factor antibodies.

DOI or PMID #

N/A

Additional Notes

Detection of Twist in the whole cell lysates of non-transfected cells might require high protein loading per gel lane.

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Summary

The Good

The antibody appears capable of detecting Twist in human carcinoma cells by western blotting

The Bad

The antibody appears to produce a number of non-specific proein bands.

The Bottom Line

The mouse monoclonal antibody sc-81417 from Santa Cruz could be used for detection of Twist in whole cell lysates of human carcinoma cells by western blotting under reducing conditions.

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