Phospho anti-Rb (S780) antibody [EPR182(N)] (ab173289)

Product name

Anti-Rb (phospho S780) antibody [EPR182(N)]
See all Rb primary antibodies
Description

Rabbit monoclonal [EPR182(N)] to Rb (phospho S780)
Host species

Rabbit
Tested applications

Suitable for: Dot blot, IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC or IP
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Rb aa 750-850 (phospho S780) (Cysteine residue). The exact sequence is proprietary.
Database link: P06400
Positive control
- Jurkat cell lysate; Human thyroid gland carcinoma and testis tissues.
General notes

A trial size is available to purchase for this antibody.

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents
This product is a recombinant rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR182(N)
Isotype

IgG
Research areas

Alternative Versions
- Anti-Rb (phospho T780) antibody [EPR182(N)] - BSA and Azide free (ab240174)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Positive Controls
- Jurkat whole cell lysate (ab30128)
- Jurkat whole cell lysate (ab7899)

Our Abpromise guarantee covers the use of ab173289 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
Dot blot		1/1000.
IHC-P		1/100.
WB		1/10000. Predicted molecular weight: 106 kDa.

Application notes

Is unsuitable for Flow Cyt,ICC or IP.

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 S780) antibody [EPR182(N)] (ab173289)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Rb (phospho S780) with ab173289 at 1/100 dilution,followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on normal human colon.

Counter stained with Hematoxylin.

Secondary antibody only control:Used PBS instead of primary antibody,secondary antibody is ab97051 at 1/500 dilution.
Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

All lanes : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/50000 dilution

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) serum starved for 44 h. Whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) serum starved for 24 h. Then 10% FBS incubated for 20 h. Whole cell lysates
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) serum starved for 24 h. Then 10% FBS incubated for 20 h. Whole cell lysates Then the membrane was 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

Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Lane 1 : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/20000 dilution
Lane 2 : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 2 : K562(Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate, serum starved for 2 days

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 106 kDa

Blocking/Dilution buffer:5% NFDM/TBST
Exposure time:1 minute
Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

All lanes : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/5000 dilution

Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 2 : Calyculin A treated K562(Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 106 kDa

Blocking/Dilution buffer:NFDM/TBST
Ab173289: anti- Rb (phospho S780);
Ab184796: anti- Rb (phospho S807);
Ab181616: anti-Rb
Dot Blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Dot blot analysis of Rb (phospho S780) peptide (Lane 1),Rb(non-phospho) peptide (Lane 2) with ab173289 at a dilution of 1/1000.ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/1000.
Blocking and dilution buffer:5% NFDM/TBST.
Exposure time:3 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Immunohistochemical analysis of paraffin-embedded Lung squamous cell carcinoma labeling Rb (phospho S780) with ab173289 at 1/100 dilution,followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on tumor cells of lung squamous cell carcinoma.
Counter stained with Hematoxylin.

Secondary antibody only control:Used PBS instead of primary antibody,secondary antibody is ab97051 at 1/500 dilution.
Western blot - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

All lanes : Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289) at 1/1000 dilution

Lane 1 : Jurkat cell lysate
Lane 2 : Jurkat cell lysate treated with Lambda Phosphatase

Lysates/proteins at 10 µg per lane.

Predicted band size: 106 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Immunohistochemical analysis of paraffin-embedded Human thyroid gland carcinoma tissue labeling Rb with ab173289 at 1/50 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Rb with ab173289 at 1/50 dilution.

Datasheet

Western blot protocols
Immunohistochemistry protocols

This product has been referenced in:

Luo Y et al. ß-catenin nuclear translocation induced by HIF-1a overexpression leads to the radioresistance of prostate cancer. Int J Oncol 52:1827-1840 (2018). Read more (PubMed: 29658569) »
Yu P et al. Androgen-independent LNCaP cells are a subline of LNCaP cells with a more aggressive phenotype and androgen suppresses their growth by inducing cell cycle arrest at the G1 phase. Int J Mol Med 40:1426-1434 (2017). Read more (PubMed: 28901378) »

See all 4 Publications for this product

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Anti-Rb (phospho S780) antibody [EPR182(N)] (ab173289)

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