Anti-Rb (phospho S249) antibody (ab4788)
- Datasheet
- References (2)
- Protocols
Overview
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Product name
Anti-Rb (phospho S249) antibody
See all Rb primary antibodies -
Description
Rabbit polyclonal to Rb (phospho S249) -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Chicken
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Immunogen
Synthetic peptide (Human) derived from a region of human Rb that contains serine 249 and threonine 252 (based on Swiss Protein database, accession number P06400). The sequence is conserved in human, mouse and rat.
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Positive control
- Jurkat cells in high growth phase.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Rb protein. The final product is generated by affinity chromatography using a Rb-derived peptide that is phosphorylated at serine 249 and threonine 252. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab4788 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Application notes
WB: 1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 106 kDa).
Can be blocked with Rb peptide - phospho S249 + T252 (phospho and non-phospho pair).
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Can be blocked with Rb peptide - phospho S249 + T252 (phospho and non-phospho pair).
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
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Function
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. -
Tissue specificity
Expressed in the retina. -
Involvement in disease
Childhood cancer retinoblastoma
Bladder cancer
Osteogenic sarcoma -
Sequence similarities
Belongs to the retinoblastoma protein (RB) family. -
Domain
The Pocket domain binds to the threonine-phosphorylated domain C, thereby preventing interaction with heterodimeric E2F/DP transcription factor complexes. -
Post-translational
modificationsPhosphorylated 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. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 386582 Chicken
- Entrez Gene: 5925 Human
- Entrez Gene: 19645 Mouse
- Entrez Gene: 24708 Rat
- Omim: 614041 Human
- SwissProt: Q90600 Chicken
- SwissProt: P06400 Human
- SwissProt: P13405 Mouse
see all -
Alternative names
- Exon 17 tumor GOS561 substitution mutation causes premature stop antibody
- GOS563 exon 17 substitution mutation causes premature stop antibody
- OSRC antibody
see all
Images
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Western blot - Anti-Rb (phospho S249) antibody (ab4788)Peptide Competition: Cell extracts prepared from Jurkat cells in high growth phase were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were incubated with 0.50µ g/mL ab4788, following prior incubation in the absence of the phosphopeptide immunogen (1), a generic phosphoserine containing peptide (2), a generic phosphothreonine containing peptide (3), the non-phosphopeptide corresponding to the phosphopeptide immunogen (4), or the presence of the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, therefore demonstrating the specificity of the ab4788 antibody for this epitope. Peptide Competition: Cell extracts prepared from Jurkat cells in high growth phase were resolved by SDS-PAGE on a 1
Datasheets and documents
References
This product has been referenced in:
- Pérez-Morales J et al. Hyper-phosphorylation of Rb S249 together with CDK5R2/p39 overexpression are associated with impaired cell adhesion and epithelial-to-mesenchymal transition: Implications as a potential lung cancer grading and staging biomarker. PLoS One 13:e0207483 (2018). Read more (PubMed: 30452490) »
- Lee YC et al. Macrophage inflammatory protein-3alpha influences growth of K562 leukemia cells in co-culture with anticancer drug-pretreated HS-5 stromal cells. Toxicology 249:116-22 (2008). WB ; Human . Read more (PubMed: 18538911) »
