ab81701 specifically stains the nuclei of BT-20 cells and primary human foreskin fibroblast (HFF) cells. It reacts with hyperphosphorylated and un (under) phosphorylated form of Rb protein. It shows no cross reaction with p107 and p130.
Human Does not react with:
Recombinant human Rb protein.
Amino acids 703-722.
Ls174T, HT29, HUVEC cells. Normal colon carcinoma. Breast carcinoma.
Abcam is committed to meeting high standards of ethical manufacturing and as such, we will be discontinuing this product, which has been generated by the ascites method, within the next year. We are sorry for any inconvenience this may cause. If you would like help finding an alternative product, please do not hesitate to contact our scientific support team.
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Constituents: 10mM PBS, pH 7.4
Concentration information loading...
Protein G purified
Purified from the ascites fluid by Protein G chromatography.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Flow Cyt: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent dilution.
IHC-P: Use at a concentration of 1 - 2 µg/ml for 30 min at room temperature. Staining of formalin fixed tissues requires boiling tissue sections in 1mM EDTA, pH 8.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes. EDTA is better than citrate for epitope unmasking.
IP (Native and denatured): Use at 2 µg/mg of lysate.
WB: Use at a concentration of 1 - 2 µg/ml for 2 hours at room temperature. Predicted molecular weight: 105 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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.
Involvement in disease
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.
Formalin fixed, paraffin embedded human breast carcinoma stained with anti Rb (form of ab81701 containing BSA and azide) at 2 µg/ml. Peroxidase conjugate and AEC chromogen were used. Note nuclear staining of tumor cells.
Western blot - Rb antibody [1F8] - BSA and Azide free (ab81701)
anti Rb (form of ab81701 containing BSA and azide) at 2 µg/ml + human cells
Predicted band size : 105 kDa Observed band size : 105 kDa