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Read our guarantee »Products:Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Rb
Anti-Rb antibody [Rb1 1F8] - Nuclear Marker
See all Rb products (41) ...
Mouse monoclonal [Rb1 1F8] to Rb - Nuclear Marker
This antibody reacts with hyperphosphorylated and un (under) phosphorylated forms of Rb protein.
WB, IP, IHC-Frmore details
Reacts with
Mouse, Human
Predicted to work with
Chicken, Chimpanzee
Rb-b-galactosidase fusion protein spanning nucleotides 1126-1973 of human Rb gel purified from bacterial lysates (R. Grand et al. 1989).
IMMCSMYG ICKVKNIDLK FKIIVTAYKD LPHAVQETF K RVLIKEEEYD SIIVFYNSVF MQRLKTNILQ YA
The epitope has been mapped between aa 703-772 of human RB1.
Tested in a panel of human cell lines and CV-1 cell line (established from monkey kidney epithelium).
Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: None
Constituents: PBS
Concentration information loading...
Protein A purified
Monoclonal
Rb1 1F8
Sp2/0-Ag14
IgG1
Cancer >> Oncoproteins/suppressors >> Tumor suppressors >> Rb family
Cancer >> Oncoproteins/suppressors >> Tumor suppressors
Epigenetics and Nuclear Signaling >> Transcription >> Cancer susceptibility >> Tumor Suppressors
Tags & Cell Markers >> Subcellular Markers >> Nucleus >> Nucleoplasm
Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Rb
Our Abpromise guarantee covers the use of ab24 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use at an assay dependent dilution. Predicted molecular weight: 105 kDa.
IP: Use at an assay dependent dilution.
IHC-Fr: Use at an assay dependent dilution.
Key regulator of entry into cell division that acts as a tumor suppressor. 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, SUV420H1 and SUV420H2, 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.
Defects in RB1 are the cause of childhood cancer retinoblastoma (RB) [MIM:180200]. RB is a congenital malignant tumor that arises from the nuclear layers of the retina. It occurs in about 1:20'000 live births and represents about 2% of childhood malignancies. It is bilateral in about 30% of cases. Although most RB appear sporadically, about 20% are transmitted as an autosomal dominant trait with incomplete penetrance. The diagnosis is usually made before the age of 2 years when strabismus or a gray to yellow reflex from pupil ('cat eye') is investigated.
Defects in RB1 are a cause of susceptibility to bladder cancer (BLC) [MIM:109800]. A malignancy originating in tissues of the urinary bladder. It often presents with multiple tumors appearing at different times and at different sites in the bladder. Most bladder cancers are transitional cell carcinomas. They begin in cells that normally make up the inner lining of the bladder. Other types of bladder cancer include squamous cell carcinoma (cancer that begins in thin, flat cells) and adenocarcinoma (cancer that begins in cells that make and release mucus and other fluids). Bladder cancer is a complex disorder with both genetic and environmental influences.
Defects in RB1 are a cause of osteogenic sarcoma (OSRC) [MIM:259500].
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 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.
N-terminus is methylated by METTL11A/NTM1 (By similarity). Monomethylated at Lys-860 by SMYD2, promoting interaction with L3MBTL1.
Nucleus.
Target information above from: UniProt accessionP06400
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Western blot - Rb antibody [Rb1 1F8] - Nuclear Marker (ab24)
![Western blot - Rb antibody [Rb1 1F8] - Nuclear Marker (ab24)](/ps/datasheet/Images/0/ab24/ab24_2.jpg)
Anti-Rb antibody [Rb1 1F8] - Nuclear Marker (ab24) at 1/1000 dilution + Human Fibroblast cell lysate(nuclear). 16 hour incubation.
Secondary
NIF 825 (provided in ECL kit) - HRP conjugated
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 105 kDa
Observed band size : 100-150 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
This image is courtesy of an Abreview submitted on 5 October 2005.
Western blot - Rb antibody [Rb1 1F8] - Nuclear Marker (ab24)
![Western blot - Rb antibody [Rb1 1F8] - Nuclear Marker (ab24)](/ps/datasheet/images/0/ab24/Rb-Primary-antibodies-ab24-1.jpg)
Predicted band size : 105 kDa
Lysates prepared from lung and kidney of VerUTR transgenic and wildtype mice were analyzed by Western Blot probed with ab24. Increased expression of Rb1 was detected in the organs from the VerUTR transgenic mice.Cells were seeded onto 6-well plates at 2×105 cells per well overnight. They were then transfected with 1 µg of VerUTR or control vector in combination with scrambled RNA or siRNA against VerUTR. Proteins were extracted 48 hours after transfection by lysing in 60 µl of lysis buffer containing protease inhibitors (150 mM NaCl, 25 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 1% Triton X-100, 8 M Urea, and 1x protease inhibitor cocktail). Tissues were disrupted in appropriate volume of lysis buffer depending on tissue weight. All samples were subjected to SDS-PAGE and then transferred to nitrocellulose membranes followed by incubating with ab24 at 1/500 dilution at 4°C overnight. The secondary antibody used was goat anti-mouse IgG at 1/2000 dilution at room temperature for 1 hour. After detection of the protein bands, the blot was stripped and re-probed with mouse monoclonal antibody against β-actin to confirm equal loading. After secondary antibody incubation, the blot was washed and detected by ECL kit in autoradiography.
Image from Lee DY et al, PLoS One 5:e13599 (2010), Fig 3.
This product has been referenced in:
See all 6 publications for this product
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![Western blot - Rb antibody [Rb1 1F8] - Nuclear Marker (ab24)](/ps/datasheet/Images/0/ab24/ab24_2.jpg)
Anti-Rb antibody [Rb1 1F8] - Nuclear Marker (ab24) at 1/1000 dilution + Human Fibroblast cell lysate(nuclear). 16 hour incubation.
Secondary
NIF 825 (provided in ECL kit) - HRP conjugated
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 105 kDa
Observed band size : 100-150 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
This image is courtesy of an Abreview submitted on 5 October 2005.
![Western blot - Rb antibody [Rb1 1F8] - Nuclear Marker (ab24)](/ps/datasheet/images/0/ab24/Rb-Primary-antibodies-ab24-1.jpg)
Predicted band size : 105 kDa
Lysates prepared from lung and kidney of VerUTR transgenic and wildtype mice were analyzed by Western Blot probed with ab24. Increased expression of Rb1 was detected in the organs from the VerUTR transgenic mice.Cells were seeded onto 6-well plates at 2×105 cells per well overnight. They were then transfected with 1 µg of VerUTR or control vector in combination with scrambled RNA or siRNA against VerUTR. Proteins were extracted 48 hours after transfection by lysing in 60 µl of lysis buffer containing protease inhibitors (150 mM NaCl, 25 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 1% Triton X-100, 8 M Urea, and 1x protease inhibitor cocktail). Tissues were disrupted in appropriate volume of lysis buffer depending on tissue weight. All samples were subjected to SDS-PAGE and then transferred to nitrocellulose membranes followed by incubating with ab24 at 1/500 dilution at 4°C overnight. The secondary antibody used was goat anti-mouse IgG at 1/2000 dilution at room temperature for 1 hour. After detection of the protein bands, the blot was stripped and re-probed with mouse monoclonal antibody against β-actin to confirm equal loading. After secondary antibody incubation, the blot was washed and detected by ECL kit in autoradiography.
Image from Lee DY et al, PLoS One 5:e13599 (2010), Fig 3.
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