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Anti-Rb (phospho S795) antibody [EPR28156-6] is a Rabbit Monoclonal antibody that is used in Rb (phospho S795) Dot Blot, Western Blot. Suitable for Human samples.

Rb (RB1) is a critical tumor suppressor that prevents transcription of E2F-regulated genes. CDK-dependent phosphorylation of RB1 induces its dissociation from E2Fs, thereby activating transcription of E2F responsive genes and triggering entry into S phase.

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Images

Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974), expandable thumbnail
  • Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974), expandable thumbnail
  • Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974), expandable thumbnail
  • Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (AB321974), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBDotICC/IFIHC-P
Human
Tested
Expected
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Synthetic peptide - Human
Not recommended
Tested
Not recommended
Not recommended
Transfected cell lysate - Human
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Transfected cell lysate - Human
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
1/1000
Notes

-

Expected
Expected

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

-

Not recommended
Not recommended

Species
Transfected cell lysate - Human, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Transfected cell lysate - Human, Mouse, Synthetic peptide - Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Transfected cell lysate - Human, Synthetic peptide - Human
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

Retinoblastoma-associated protein, p105-Rb, p110-RB1, pRb, pp110, Rb, RB1

Recommended products

Anti-Rb (phospho S795) antibody [EPR28156-6] is a Rabbit Monoclonal antibody that is used in Rb (phospho S795) Dot Blot, Western Blot. Suitable for Human samples.

Rb (RB1) is a critical tumor suppressor that prevents transcription of E2F-regulated genes. CDK-dependent phosphorylation of RB1 induces its dissociation from E2Fs, thereby activating transcription of E2F responsive genes and triggering entry into S phase.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR28156-6
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

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.

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.

4 product images

  • Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974), expandable thumbnail

    Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    In Western blot, Anti-Rb antibody [EPR17512-291] (Anti-Rb antibody [EPR17512] ab181616) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

    Lane 1: 293T cells transfected with a human Rb expression vector containing a His-tag, whole cell lysate at 20 µg

    Lane 2: 293T cells transfected with a human Rb (S795A mutation) expression vector containing a His-tag, whole cell lysate at 20 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

    Observed band size: 106 kDa, 124 kDa

    Exposure time: 1s

  • Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974), expandable thumbnail

    Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

    This blot was developed using a high sensitivity ECL substrate.

    The identity of the bands at approximately 40 kDa, 60 kDa and 280 kDa (in lanes 1-2) are unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-Rb antibody [EPR17512-291] (Anti-Rb antibody [EPR17512] ab181616) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

    Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) serum starvation for 48h, whole cell lysate (untreated membrane) at 50 µg

    Lane 2: MCF7 serum starvation for 48h, then treated with 10% FBS for 20h, whole cell lysate (untreated membrane) at 50 µg

    Lane 3: MCF7 serum starvation for 48h, then treated with 10% FBS for 20h, whole cell lysate (alkaline phosphatase treated membrane) at 50 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

    Developed using the ECL technique.

    Observed band size: 106 kDa, 36 kDa

    Exposure time: 92s

  • Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974), expandable thumbnail

    Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-Rb antibody [EPR17512-291] (Anti-Rb antibody [EPR17512] ab181616) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

    Lane 1: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) serum starvation for 48h, whole cell lysate (untreated membrane) at 50 µg

    Lane 2: Jurkat serum starvation for 48h, then treated with 10% FBS for 20h, whole cell lysate (untreated membrane) at 50 µg

    Lane 3: Jurkat serum starvation for 48h, then treated with 10% FBS for 20h, whole cell lysate (alkaline phosphatase treated membrane) at 50 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

    Observed band size: 106 kDa, 36 kDa

    Exposure time: 15s

  • Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974), expandable thumbnail

    Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974)

    Dot blot analysis of Rb using ab321974 at 1:1000 (0.504 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

    Lane1: Human Rb (phospho S795) peptide a

    Lane2: Human Rb (phospho S795) peptide b

    Lane3: Human Rb non-phospho peptide

    Lane4: Human Rb (phospho S794) peptide

    Lane5: Mouse Rb (phospho S788) peptide

    Exposure time: 180 seconds.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    All lanes: Dot Blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (ab321974) at 1/1000 dilution

    Lane 1: Human Rb (phospho S795) peptide a

    Lane 2: Human Rb (phospho S795) peptide b

    Lane 3: Human Rb non-phospho peptide

    Lane 4: Human Rb (phospho S794) peptide

    Lane 5: Mouse Rb (phospho S788) peptide

    Secondary

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

    Exposure time: 180s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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