Western Blot of HeLa Whole-Cell Lysates Using AND-1 Antibody

February 22, 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:

BioLegend

Product Name:

Purified anti-AND-1 (WDHD1) Antibody

Catalog Number:

630301

Our aim was to examine the levels of AND-1 in a scramble and target siRNA transfected HeLa cells. Whole-cell lysates were prepared and loaded on SDS-PAGE and transferred to PVDF membrane and probed with AND-1 antibody from BioLegend.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa whole-cell lysate

Primary Incubation

1:3000, Overnight at 4°C

Blocking Agent

5% skimmed milk, 1 hr at room temperature

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 observed a single specific band corresponding to AND-1 at 126 kDa in HeLa whole-cell lysates and the levels of AND-1 was unchanged in both scramble and target siRNA transfected cells.

DOI or PMID #

29855479

Additional Notes

N/A

Related Categories

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Summary

The Good

Good antibody for performing western blots

The Bad

N/A

The Bottom Line

A single specific band was observed in HeLa cells at 126 kDa corresponding to molecular weight of AND-1. Easy to optimize and doesn't give non-specific bands by western blot.

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