Anti-6X His tag® antibody - ChIP Grade (ab9108)

  ChIP - 6X His tag antibody - ChIP Grade (ab9108)

A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a Myc or His - tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach).

  Western blot - 6X His tag® antibody - ChIP Grade (ab9108)

Use of ab9108 in western blotting at 1:1000 against E.coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 11 different epitope tag (ab5395). Lane 1: 2 µl lysate Lane 2: 1 µl lysate Lane 3: 0.5 µl lysate Lane 4: 0.25 µl lysate

  Western blot - 6X His tag antibody - ChIP Grade (ab9108)

All lanes : Anti-6X His tag® antibody - ChIP Grade (ab9108) at 1/1000 dilution

Lane 1 : 10ug non-transformed lysate
Lane 2 : 10ug non-transformed lysate + 1mM IPTG
Lane 3 : 10ug E.coli Lysate harbouring His-tag-RelA NTD, non-induced
Lane 4 : 10ug Induction of His-tag RelA NTD 1 mM IPTG
Lane 5 : 10ug Induction of His-tag RelA NTD 1 mM IPTG

Secondary
HRP conjugated goat anti-rabbit IgG
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 37 kDa (why is the actual band size different from the predicted?)
Additional bands at : 12 kDa (possible non-specific binding).

Exposure time : 1 minute

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