First purified by protein G affinity chromatography, then the antibody fraction is peptide affinity purified in a 2 step procedure with the control and phosphorylated peptides. The phospho specific antibody is eluted with high and low pH buffers and neutralized immediately, followed by dialysis against PBS.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/100.
Key regulator of entry into cell division that acts as a tumor suppressor. Promotes G0-G1 transition when phosphorylated by CDK3/cyclin-C. Acts as a transcription repressor of E2F1 target genes. The underphosphorylated, active form of RB1 interacts with E2F1 and represses its transcription activity, leading to cell cycle arrest. Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation. Recruits and targets histone methyltransferases SUV39H1, KMT5B and KMT5C, leading to epigenetic transcriptional repression. Controls histone H4 'Lys-20' trimethylation. Inhibits the intrinsic kinase activity of TAF1. Mediates transcriptional repression by SMARCA4/BRG1 by recruiting a histone deacetylase (HDAC) complex to the c-FOS promoter. In resting neurons, transcription of the c-FOS promoter is inhibited by BRG1-dependent recruitment of a phospho-RB1-HDAC1 repressor complex. Upon calcium influx, RB1 is dephosphorylated by calcineurin, which leads to release of the repressor complex (By similarity). In case of viral infections, interactions with SV40 large T antigen, HPV E7 protein or adenovirus E1A protein induce the disassembly of RB1-E2F1 complex thereby disrupting RB1's activity.
Expressed in the retina.
Childhood cancer retinoblastoma Bladder cancer Osteogenic sarcoma
Belongs to the retinoblastoma protein (RB) family.
The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes.
Phosphorylated by CDK6 and CDK4, and subsequently by CDK2 at Ser-567 in G1, thereby releasing E2F1 which is then able to activate cell growth. Dephosphorylated at the late M phase. SV40 large T antigen, HPV E7 and adenovirus E1A bind to the underphosphorylated, active form of pRb. Phosphorylation at Thr-821 and Thr-826 promotes interaction between the C-terminal domain C and the Pocket domain, and thereby inhibits interactions with heterodimeric E2F/DP transcription factor complexes. Dephosphorylated at Ser-795 by calcineruin upon calcium stimulation. CDK3/cyclin-C-mediated phosphorylation at Ser-807 and Ser-811 is required for G0-G1 transition. Phosphorylated by CDK1 and CDK2 upon TGFB1-mediated apoptosis. N-terminus is methylated by METTL11A/NTM1 (By similarity). Monomethylation at Lys-810 by SMYD2 enhances phosphorylation at Ser-807 and Ser-811, and promotes cell cycle progression. Monomethylation at Lys-860 by SMYD2 promotes interaction with L3MBTL1. Acetylation at Lys-873 and Lys-874 regulates subcellular localization, at least during keratinocytes differentiation.
ab60025 at 1/50 dilution staining formalin fixed, paraffin embedded human breast cancer tissue. Peroxidase conjugated to the secondary antibody followed by AEC staining.
Immunocytochemistry/ Immunofluorescence - Anti-Rb (phospho S608) antibody (ab60025)This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab60025 (1/200) staining Rb (phospho S608) in assynchonous Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X-100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.
Heller EA et al. Locus-specific epigenetic remodeling controls addiction- and depression-related behaviors. Nat Neurosci17:1720-7 (2014).
Read more (PubMed: 25347353) »
Shirazi Fard S et al. Heterogenic final cell cycle by chicken retinal Lim1 horizontal progenitor cells leads to heteroploid cells with a remaining replicated genome. PLoS One8:e59133 (2013).
Read more (PubMed: 23527113) »