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Rabbit Recombinant Monoclonal RB phospho T356 antibody. Carrier free. Suitable for IHC-P, IP, Dot, WB and reacts with Human, Synthetic peptide samples.

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Images

Immunoprecipitation - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (AB247461), expandable thumbnail
  • Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (AB247461), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (AB247461), expandable thumbnail
  • Dot Blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (AB247461), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (AB247461), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPFlow CytDotWBICC/IF
Human
Tested
Tested
Not recommended
Expected
Tested
Not recommended
Synthetic peptide
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Synthetic peptide
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide
Dilution info
-
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Synthetic peptide
Dilution info
-
Notes

-

Target data

Function

Tumor suppressor that is a key regulator of the G1/S transition of the cell cycle (PubMed:10499802). The hypophosphorylated form binds transcription regulators of the E2F family, preventing transcription of E2F-responsive genes (PubMed:10499802). Both physically blocks E2Fs transactivating domain and recruits chromatin-modifying enzymes that actively repress transcription (PubMed:10499802). Cyclin and CDK-dependent phosphorylation of RB1 induces its dissociation from E2Fs, thereby activating transcription of E2F responsive genes and triggering entry into S phase (PubMed:10499802). RB1 also promotes the G0-G1 transition upon phosphorylation and activation by CDK3/cyclin-C (PubMed:15084261). 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). (Microbial infection) 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.

Alternative names

Recommended products

Rabbit Recombinant Monoclonal RB phospho T356 antibody. Carrier free. Suitable for IHC-P, IP, Dot, WB and reacts with Human, Synthetic peptide samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR2153AY
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab247461 is the carrier-free version of Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298.

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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 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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Retinoblastoma protein (Rb) also known as pRb is an important regulatory protein with a molecular weight of approximately 110 kDa. It is mainly expressed in the nucleus of cells. Rb functions as a tumor suppressor by controlling the cell cycle progression from G1 to S phase. Rb becomes active when it is dephosphorylated allowing it to bind and inhibit E2F transcription factors consequently preventing the transcription of genes essential for S phase entry.

Biological function summary

Retinoblastoma protein influences cellular proliferation differentiation and apoptosis. It acts within a larger protein complex modulating various cellular responses. When functional Rb halts uncontrolled cell division important for maintaining normal tissue homeostasis. In its phosphorylated state known as phospho-Rb or phospho-Rb E182 it loses its regulatory capabilities which can lead to unrestrained cell cycle progression.

Pathways

Several involve the retinoblastoma protein. One key pathway is the p53 pathway which Rb interacts with to influence cellular outcomes like cell-cycle arrest and apoptosis. Rb cooperates with proteins like p21 to implement these processes. Additionally it is involved in the cyclin D-dependent kinase 4 (CDK4) pathway which modulates its phosphorylation state influencing binding interactions and cell cycle control.

Associated diseases and disorders

Dysregulation of retinoblastoma protein is commonly associated with cancer particularly retinoblastoma and breast cancer. In retinoblastoma mutations in the Rb gene directly lead to uncontrolled cell proliferation due to the absence of functional Rb protein. Additionally Rb's connection to the E2F family of transcription factors can become disrupted contributing to oncogenesis in other cancers.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunoprecipitation - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461), expandable thumbnail

    Immunoprecipitation - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.
    Purified Anti-Rb (phospho T356) antibody [EPR2153AY] 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 (+): Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298 + Jurkat whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298 in Jurkat whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (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)

    All lanes: Immunoprecipitation - Anti-Rb (phospho T356) antibody [EPR2153AY] (Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298)

    Predicted band size: 106 kDa

  • Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461), expandable thumbnail

    Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.

    All lanes: Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] (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. at 15 µg

    Lane 2: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate. at 15 µg

    Lane 3: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate. Membrane incubated with phosphatase. at 15 µg

    Secondary

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

    Predicted band size: 106 kDa

    Observed band size: 106 kDa

    Exposure time: 1min

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298 showing positive staining in Endometrial carcinoma tissue.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Dot Blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461), expandable thumbnail

    Dot Blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.Dot blot analysis of Rb (phospho T356) phospho peptide (Lane 1), Rb Non-phospho peptide (Lane 2), labelling Rb (phospho T356) with Anti-Rb (phospho T356) antibody [EPR2153AY] 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] - BSA and Azide free (ab247461), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.Anti-Rb (phospho T356) antibody [EPR2153AY] 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] - BSA and Azide free (ab247461), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.Anti-Rb (phospho T356) antibody [EPR2153AY] 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] - BSA and Azide free (ab247461), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.Anti-Rb (phospho T356) antibody [EPR2153AY] 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] - BSA and Azide free (ab247461), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Anti-Rb (phospho T356) antibody [EPR2153AY] 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] - BSA and Azide free (ab247461), expandable thumbnail

    Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461)

    This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.

    All lanes: Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] - BSA and Azide free (ab247461) at 1/2000 dilution

    Lane 1: Jurkat cell lysate at 10 µg

    Lane 2: Jurkat cell lysate treated with FBS at 10 µg

    Secondary

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

    Predicted band size: 106 kDa

    Observed band size: 106 kDa

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Product protocols

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