The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
1/2000 - 1/10000. Detects a band of approximately 324 kDa (predicted molecular weight: 324 kDa).
Several other bands.
Use at 2-5 µg/mg of lysate.
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
RelevanceBPTF (bromodomain and PHD domain transcription factor) is the largest subunit of the ATP-dependent chromatin-remodelling complex, NURF (nucleosome remodelling factor). NURF catalyses ATP-dependent nucleosome sliding and facilitates transcription. BPTF recognises histone H3 tails that are tri-methylated at K4, which marks the transcriptional start site of the vast majority of transcriptionally active genes. BPTF also exhibits some binding to H3 di-methylated at K4. BPTF plays a key role in the development of early mouse embryos, possibly through regulation of the Smad pathway of transcription factors. While BPTF is expressed in low levels in the adult brain and spinal cord, it is expressed in higher levels in the brain in neurodegenerative diseases. It is present in a subset of amyloid-containing plaques in the brains of patients suffering from Alzheimer's disease.
Immunoprecipitation/ Western Blot of FALZ / BPTF.
Lane 1: ab72036 at 3µg/mg whole cell lysate for IP, and 0.1 µg/ml for subsequent WB.
Lane 2: Control IgG.
Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded.
Detection: Chemiluminescence with an exposure time of 1 second.
ICC/IF image of ab72036 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab72036, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-BPTF / FALZ antibody (ab72036)
This product has been referenced in:
Mulder KW et al. Diverse epigenetic strategies interact to control epidermal differentiation. Nat Cell Biol14:753-63 (2012).
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