ChIPing with TAGs

By Dr. Hugh Spotswood *

Antibodies to HA, Myc, His, T7 and V5 - all ChIP Grade!

Drosophila and yeast chromatin biology in particular have benefited from the use of tags because of the relative ease in which their genomes are manipulated. Epitope tagging permits the analysis of the function, interaction and the sub-cellular localisation of a protein target without the cost and time delay of raising a new antibody against the endogenous target. It also permits the analysis of proteins that would be otherwise too difficult to purify due to the similarity they share with other paralogues.

At Abcam we now have a panel of antibodies against the major epitope tags; HA, Myc, His, T7 and V5 for use in chromatin immunoprecipitation (ChIP).

ChIP Grade Tag Antibodies


AbID

Target

Epitope Tag

9110

Rabbit polyclonal to HA Tag

HA

1424

Mouse monoclonal [4C12] to HA tag

HA

9132

Goat polyclonal to Myc tag

Myc

9108

Rabbit polyclonal to 6xHis tag

His

9138

Goat polyclonal to T7 tag

T7

15828

Rabbit polyclonal to V5 tag

V5


We have validated our antibodies against the epitope tags using a human cell line stably transfected with the Gal4 upstream activation sequence. Cells were transiently transfected with epitope tagged Gal4 DNA Binding Domain constructs. Our epitope tagged antibodies were assayed for their capacity to chromatin immunoprecipitate by performing ChIP in parallel using antibodies against Gal4 (ChIP grade; ab1396) and the respective epitope tag. The enrichment across the Gal4 upstream activation sequence was assayed by real time PCR.

The results (see figure below for example) showed that the antibodies that were confirmed as working in ChIP demonstrated a similar level of enrichment at the Gal4 upstream activation sequence using anti-Gal4 antibody (ab1396) to the epitope tagged (anti-T7 and anti-V5) antibodies.

ChIP with anti-T7 tag antibody (ab9138)

ab9138_resize

ChIP with anti-V5 antibody (ab15828)

ab15828_resize

Abcam now has a comprehensive panel of ChIP grade antibodies against the major epitope tags for use in a broad range of experimental systems. At Abcam we make a promise (our Abpromise) to our customers that our antibodies behave as detailed on our datasheets - and, where ChIP-Grade is noted, that means we guarantee that all of our new epitope tagged antibodies work by chromatin immunoprecipitation too!

Useful links:


Hugh Spotswood, Senior Technical Support Specialist, Abcam plc

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