Product Images
Immunocytochemistry Analysis:
Representative lot data:
Immunofluorescent analysis of HeLa cells using Anti-phospho-Rb (Thr821), clone ...
Western Blot Analysis:
Representative lot data:
Phosphatase untreated and treated Jurkat cell lysates were probed Anti-phospho-Rb (Thr...
Anti-phospho-Retinoblastoma (Thr821), clone E231, Rabbit Monoclonal
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, IC, IP | Rabbit | Diluted Culture Supernatant | Monoclonal Antibody |
Description:
Anti-phospho-Retinoblastoma (Thr821), clone E231, Rabbit Monoclonal
Molecular Weight:
~110 observed
Epitope:
Phosphorylated Thr821
Immunogen:
Synthetic phospho-peptide corresponding to human Rb phosphorylated at Thr821.
Modifications:
Phosphorylation
Clone:
E231
Isotype:
IgG
Background Information:
Retinoblastoma tumor suppressor protein (Rb) is associated with DNA binding activity and is thought to play a significant role in controlling cell cycle progression during tumor growth (1,2). Cell cycle-dependent phosphorylation by cdk’s inhibits Rb binding, thus allowing cell cycle progression (3). Rb inactivation and cell cycle progression likely requires first phosphorylation by cyclin Dcdk4/ 6 followed by cyclin E-cdk2 phosphorylation (4).
Species Reactivity:
Human
Species Reactivity Note:
Demonstrated to react with human.
Application Notes:
Immunoprecipitation Analysis: 1:20 dilution from a representative lot was used in IP.
Immunocytochemistry Analysis: 1:100-1:250 dilution from a representative lot detected Rb in HeLa cells.
Immunocytochemistry Analysis: 1:100-1:250 dilution from a representative lot detected Rb in HeLa cells.
Control:
Phosphatase untreated and treated Jurkat cell lysates
Quality Assurance:
Evaluated by Western Blot in Phosphatase untreated and treated Jurkat cell lysates.
Western Blot Analysis: 1:1,000 dilution of this antibody detected Rb in Phosphatase untreated and treated Jurkat cell lysates.
Western Blot Analysis: 1:1,000 dilution of this antibody detected Rb in Phosphatase untreated and treated Jurkat cell lysates.
Purification Method:
Unpurified
Presentation:
Rabbit Monoclonal in buffer containing 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl containing 40% Glycerol, 0.01% sodium azide and 0.05% BSA.
Storage Conditions:
Stable for 1 year at -20ºC from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below
-20°C may cause glycerol containing Solutions to become frozen during storage.
-20°C may cause glycerol containing Solutions to become frozen during storage.
UniProt Number:
Gene Symbol:
- retinoblastoma 1
- RB
- p105-Rb
- pp110
- pRb
- osteosarcoma
- OSRC
- Rb
Alternate Names:
- retinoblastoma suspectibility protein
- retinoblastoma-associated protein
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
- Western Blotting
- Immunocytochemistry
- Immunoprecipitation
UniProt Summary:
FUNCTION: 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.
SUBUNIT STRUCTURE: Interacts with ATAD5. Interacts with PRMT2 By similarity. The hypophosphorylated form interacts with and sequesters the E2F1 transcription factor. Interacts with heterodimeric E2F/DP transcription factor complexes containing TFDP1 and either E2F1, E2F3, E2F4 or E2F5, or TFDP2 and E2F4. The unphosphorylated form interacts with EID1, ARID3B, KDM5A, SUV39H1, MJD2A/JHDM3A and THOC1. Interacts with the N-terminal domain of TAF1. Interacts with AATF, DNMT1, LIN9, LMNA, SUV420H1, SUV420H2, PELP1 and TMPO-alpha. May interact with NDC80. Interacts with GRIP1 and UBR4. Interacts with ARID4A and KDM5B. Interacts with E4F1 and LIMD1. Interacts with SMARCA4/BRG1 AND HDAC1 By similarity. Interacts with adenovirus E1A protein, HPV E7 protein and SV40 large T antigen. Interacts with PSMA3 and USP4. Interacts (when methylated at Lys-860) with L3MBTL1.
SUBCELLULAR LOCATION: Nucleus.
TISSUE SPECIFICITY: Expressed in the retina.
DOMAIN: The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes.
PTM: 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.
INVOLVEMENT IN DISEASE: 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].
SEQUENCE SIMILARITIES: Belongs to the retinoblastoma protein (RB) family.
SUBUNIT STRUCTURE: Interacts with ATAD5. Interacts with PRMT2 By similarity. The hypophosphorylated form interacts with and sequesters the E2F1 transcription factor. Interacts with heterodimeric E2F/DP transcription factor complexes containing TFDP1 and either E2F1, E2F3, E2F4 or E2F5, or TFDP2 and E2F4. The unphosphorylated form interacts with EID1, ARID3B, KDM5A, SUV39H1, MJD2A/JHDM3A and THOC1. Interacts with the N-terminal domain of TAF1. Interacts with AATF, DNMT1, LIN9, LMNA, SUV420H1, SUV420H2, PELP1 and TMPO-alpha. May interact with NDC80. Interacts with GRIP1 and UBR4. Interacts with ARID4A and KDM5B. Interacts with E4F1 and LIMD1. Interacts with SMARCA4/BRG1 AND HDAC1 By similarity. Interacts with adenovirus E1A protein, HPV E7 protein and SV40 large T antigen. Interacts with PSMA3 and USP4. Interacts (when methylated at Lys-860) with L3MBTL1.
SUBCELLULAR LOCATION: Nucleus.
TISSUE SPECIFICITY: Expressed in the retina.
DOMAIN: The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes.
PTM: 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.
INVOLVEMENT IN DISEASE: 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].
SEQUENCE SIMILARITIES: Belongs to the retinoblastoma protein (RB) family.
Protein/Isoform Description:
Rb is a tumor suppressor protein with a pocket domain, which binds to HDAC1. It also binds to the E2F transcription factors, and the complex regulates E2F-responsive promoters of genes involved in DNA replication. Rb-associated HDAC1 deacetylates histones at E2F-responsive promoters, thereby repressing their transcription. Cdks phosphorylate Rb at the G1/S boundary, causing it to dissociate from E2F. HDAC1 dissociates from E2F along with Rb, and E2F is then able to stimulate transcription of genes important for DNA replication.
Product Name:
Anti-phospho-Retinoblastoma (Thr821), clone E231, Rabbit Monoclonal
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 µL
Format:
Diluted Culture Supernatant
Host:
Rabbit