Western blot analysis was performed on nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), MDA-MB-231 (Lane 3), HEK 293 (Lane 4), SW480 (Lane 5), HT-29 (Lane 6), SH-SY5Y (Lane 7), F9 (Lane 8) and T47D (Lane 9). The blot was probed with Rabbit Anti-RbAp46 Polyclonal Antibody (Product # PA1-1868, 2 µg/ml) and detected by chemiluminescence. A 50 kDa band corresponding to RbAp46 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary antibody following blocking with 5 % skimmed milk. This is followed by incubating the membrane with Poly-HRP Streptavidin (Product# N200, 1:10,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) A S K E M F E D T V E E(13) C of human RbAp46.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1.1-868 detects retinoblastoma protein (Rb) associated protein 46 from human cells.
PA1.1-868 has been successfully used in Western blot, ICC/IF and IHC applications. By Western blot, this antibody detects an ~46 kDa protein representing RbAp46 Biotin from HeLa cell lysate
The PA1.1-868 immunogen is a synthetic peptide corresponding to residues M(1) A S K E M F E D T V E E(13) C of human RbAp46. This sequence is completely conserved between human and mouse. PA1.1-868 immunizing peptide (Cat. # PEP-083) is available for use in neutralization and control experiments.
Retinoblastoma protein (Rb) associated protein 46 (RbAp46) is a nuclear protein that has been shown to interact directly with retinoblastoma protein and is a component of the mSin3 corepressor complex as are the histone deacetylase proteins (HDACs). RbAp46 shares about 75% sequence identity with the closely related RbAp48, and appears to be functionally related to the yeast MSI1 protein in that overexpression of RbAp46 suppresses heat shock sensitivity in a RAS pathway dependent manner. RbAp46 has also been demonstrated to be up-regulated by the Wilms and quote; tumor suppressor gene product, WT1. Additionally, cells transfected with RbAp46 have shown strong inhibition of growth.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.