Phospho Rb (T356) antibody [EPR2153AY] Recombinant (ab76298)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2153AY] to Rb (phospho T356)
Suitable for: WB, IP, IHC-P, Dot blot
Reacts with: Human

Product name

Anti-Rb (phospho T356) antibody [EPR2153AY]
See all Rb primary antibodies
Description

Rabbit monoclonal [EPR2153AY] to Rb (phospho T356)
Host species

Rabbit
Specificity

The antibody only detects Rb phosphorylated on Threonine 356.

Tested Applications & Species

Application	Species
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Jurkat cell lysate treated with FBS Human breast carcinoma tissue HeLa cells IP: Jurkat whole cell lysate.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR2153AY
Isotype

IgG
Research areas

Alternative Versions
- Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Rb protein (ab83205)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab76298 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
IHC-P	Human
IP	Human
WB	Human

Application	Abreviews	Notes
WB		1/2000. Detects a band of approximately 110 kDa (predicted molecular weight: 106 kDa).
IP		1/20.
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Dot blot		Use at an assay dependent concentration.

Notes
WB 1/2000. Detects a band of approximately 110 kDa (predicted molecular weight: 106 kDa).
IP 1/20.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Dot blot Use at an assay dependent concentration.

Application notes

Is unsuitable for Flow Cyt or ICC/IF.

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
modifications

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.
Cellular localization

Nucleus.
Target information above from: UniProt accession P06400 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5925 Human
- Omim: 614041 Human
- SwissProt: P06400 Human
- Unigene: 408528 Human
Alternative names
- Exon 17 tumor GOS561 substitution mutation causes premature stop antibody
- GOS563 exon 17 substitution mutation causes premature stop antibody
- OSRC antibody
- Osteosarcoma antibody
- p105-Rb antibody
- P105RB antibody
- PP105 antibody
- pp110 antibody
- PPP1R130 antibody
- pRb antibody
- Prepro retinoblastoma associated protein antibody
- Protein phosphatase 1 regulatory subunit 130 antibody
- Rb antibody
- RB transcriptional corepressor 1 antibody
- RB_HUMAN antibody
- RB1 antibody
- RB1 gene antibody
- Retinoblastoma 1 antibody
- Retinoblastoma suspectibility protein antibody
- Retinoblastoma-associated protein antibody
see all

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

ab76298 showing positive staining in Endometrial carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

All lanes : Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298) at 1/4000 dilution

Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) 2 day serum starved whole cell lysate.
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate.
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate. Membrane incubated with phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 106 kDa
Observed band size: 106 kDa

Exposure time: 1 minute
Immunoprecipitation - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

Purified ab76298 at 1/20 dilution (0.5µg) immunoprecipitating Rb in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab76298 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76298 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 110 kDa
Cleavage fragments were observed as described in literature. (PMID: 11420704)
Dot Blot - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

Dot blot analysis of Rb (phospho T356) phospho peptide (Lane 1), Rb Non-phospho peptide (Lane 2), labelling Rb (phospho T356) with ab76298 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

ab76298 showing positive staining in Lung adenocarcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

ab76298 showing positive staining in Ovarian carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

ab76298 showing positive staining in Normal tonsil tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab76298 at 1/100-1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

All lanes : Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298) at 1/2000 dilution

Lane 1 : Jurkat cell lysate
Lane 2 : Jurkat cell lysate treated with FBS

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 106 kDa
Observed band size: 106 kDa
Anti-Rb (phospho T356) antibody [EPR2153AY] (ab76298)

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet

Publishing research using ab76298? Please let us know so that we can cite the reference in this datasheet.

ab76298 has been referenced in 5 publications.

Dai L et al. SETD4 Regulates Cell Quiescence and Catalyzes the Trimethylation of H4K20 during Diapause Formation in Artemia. Mol Cell Biol 37:N/A (2017). PubMed: 28031330
Attiyeh EF et al. Pharmacodynamic and genomic markers associated with response to the XPO1/CRM1 inhibitor selinexor (KPT-330): A report from the pediatric preclinical testing program. Pediatr Blood Cancer 63:276-86 (2016). PubMed: 26398108
Beck TN et al. EGFR and RB1 as Dual Biomarkers in HPV-Negative Head and Neck Cancer. Mol Cancer Ther 15:2486-2497 (2016). PubMed: 27507850
Beck TN et al. Phospho-T356RB1 predicts survival in HPV-negative squamous cell carcinoma of the head and neck. Oncotarget 6:18863-74 (2015). IHC-P ; Human . PubMed: 26265441
Li S et al. MicroRNA-490-5p inhibits proliferation of bladder cancer by targeting c-Fos. Biochem Biophys Res Commun 441:976-81 (2013). WB ; Human . PubMed: 24220339

Customer reviews and Q&As

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (5)

Customer reviews and Q&As

Post-translational
modifications