Western blot analysis of extracts from HeLa cells, mock transfected (-) or transfected with a construct expressing full-length human Tip60 (hTip60; +), using Tip60 Antibody.Learn more about how we get our images.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Tip60 Antibody recognizes transfected levels of total Tip60 protein.
Mouse, Monkey, Bovine, Guinea Pig, Horse
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding His60 of human Tip60 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Tip60 is a member of the MYST (MOZ, YBF2, SAS2 and Tip60) family of histone acetyltransferases and plays a role in a variety of cellular processes such as transcriptional regulation, DNA repair, and apoptosis (1,2). Tip60 exists as part of a multi-subunit complex that includes proteins such as TRRAP, p400, Reptin, and Pontin (3,4). Tip60 plays important roles in double-stranded DNA break (DSB) repair. Tip60 is required for the activation of the ATM kinase in response to DSBs, as well as acetylation of histones H4 and H2A.X at DSBs to facilitate DNA repair (1,2,5-7). In addition, Tip60 dependent acetylation at Lys120 of p53 within the DNA binding domain is required for the induction of apoptosis upon DNA damage (8,9). Tip60 is involved in a number of transcriptional regulation pathways driven by factors such as nuclear receptors and β-catenin (10-13). The Tip60 complex has been shown to be important for mouse embryonic stem cell self-renewal by regulating transcription of developmental regulators that are controlled by Nanog (14). GSK3 (glycogen synthase kinase-3) mediated phosphorylation at Ser86 of Tip60 promotes Tip60 acetylation and subsequent stimulation of the required autophagy protein ULK1 (15).
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|12058S||100 µl (10 western blots)||$ 255.0|