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.
Histone-binding component of NURF (nucleosome-remodeling factor), a complex which catalyzes ATP-dependent nucleosome sliding and facilitates transcription of chromatin. Specifically recognizes H3 tails trimethylated on 'Lys-4' (H3K4me3), which mark transcription start sites of virtually all active genes. May also regulate transcription through direct binding to DNA or transcription factors.
Ubiquitously expressed, with highest levels in testis. Present in kidney, liver and brain. In the brain, highest levels are found in motor cortex (at protein level).
Belongs to the PBTF family. Contains 1 bromo domain. Contains 1 DDT domain. Contains 2 PHD-type zinc fingers.
Abundantly expressed in the fetal brain. Present throughout the gray and white matter of the developing spinal cord at 18-22 gestational weeks. Expressed at low levels in adult brain and spinal cord and reexpressed in neurodegenerative diseases (at protein level).
The second PHD-type zinc finger mediates binding to histone H3K4Me3. Has specificity for trimethyllysine; introducing a mutation in the Tyr-2876 residue can induce binding to dimethyllysine.
Phosphorylation enhances DNA-binding. Highly susceptible to proteolysis.
Cytoplasm. Nucleus. In brains of Alzheimer disease patients, present in a subset of amyloid-containing plaques.
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.