This antibody gave a positive signal in the following Lysates:
HeLa Whole Cell Lysate
Rat Lung Tissue
Human Lung Tissue
Mouse Spleen Tissue
This antibody gave a positive result in IHC in the following FFPE tissue: Human normal testis.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.1 - 1 µg/ml.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 48 kDa).
Core histone-binding subunit that may target chromatin assembly factors, chromatin remodeling factors and histone deacetylases to their histone substrates in a manner that is regulated by nucleosomal DNA. Component of several complexes which regulate chromatin metabolism. These include the chromatin assembly factor 1 (CAF-1) complex, which is required for chromatin assembly following DNA replication and DNA repair; the core histone deacetylase (HDAC) complex, which promotes histone deacetylation and consequent transcriptional repression; the nucleosome remodeling and histone deacetylase complex (the NuRD complex), which promotes transcriptional repression by histone deacetylation and nucleosome remodeling; the PRC2/EED-EZH2 complex, which promotes repression of homeotic genes during development; and the NURF (nucleosome remodeling factor) complex.
Belongs to the WD repeat RBAP46/RBAP48/MSI1 family. Contains 6 WD repeats.
IHC image of RbAp48 staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab47456, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-RbAp48 antibody (ab47456)This image is courtesy of an anonymous Abreview
All lanes : Anti-RbAp48 antibody (ab47456) at 1/2500 dilution
Lane 1 : Mouse MEL (erythroleukemia) cell lysate Lane 2 : Human K562 cell lysate
Lysates/proteins at 25 µg per lane.
Secondary All lanes : Donkey Anti-Rabbit IgG H&L (HRP) (ab6802) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 48 kDa Observed band size: 48 kDa
All lanes : Anti-RbAp48 antibody (ab47456) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Lung (Rat) Tissue Lysate Lane 3 : Human lung normal tissue lysate Lane 4 :Mouse spleen tissue lysate - total protein (ab29293)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed at 1/3000 dilution
ICC/IF image of ab47456 stained HeLa cells. The cells were 4% PFA fixed (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 (ab47456, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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). This antibody also gave a positive result in 4% pfa fixed (10 min) HepG2, MCF-7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2, MCF-7 cells at 1µg/ml.