Overview

  • Product name
    Anti-Rb antibody [Rb1 1F8]
    See all Rb primary antibodies
  • Description
    Mouse monoclonal [Rb1 1F8] to Rb
  • Specificity
    This antibody reacts with hyperphosphorylated and un (under) phosphorylated forms of Rb protein.
  • Tested applications
    Suitable for: WB, IP, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chicken, Chimpanzee
  • Immunogen

    Rb-b-galactosidase fusion protein spanning nucleotides 1126-1973 of human Rb gel purified from bacterial lysates (R. Grand et al. 1989).

  • Epitope
    The epitope has been mapped between aa 703-772 of human RB1.
  • Positive control
    • Tested in a panel of human cell lines and CV-1 cell line (established from monkey kidney epithelium).

Properties

Applications

Our Abpromise guarantee covers the use of ab24 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 Use at an assay dependent concentration. Predicted molecular weight: 105 kDa.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Target

  • 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.
  • Information by UniProt
  • Database links
  • 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

Anti-Rb antibody [Rb1 1F8] images

  • Anti-Rb antibody [Rb1 1F8] (ab24) at 1/1000 dilution + Human Fibroblast cell lysate(nuclear). 16 hour incubation.

    Secondary
    NIF 825 (provided in ECL kit) - HRP conjugated
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 105 kDa
    Observed band size : 100-150 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an Abreview submitted on 5 October 2005.

    See Abreview



  • Predicted band size : 105 kDa

    Image from Lee DY et al, PLoS One 5:e13599 (2010), Fig 3.

    Lysates prepared from lung and kidney of VerUTR transgenic and wildtype mice were analyzed by Western Blot probed with ab24. Increased expression of Rb1 was detected in the organs from the VerUTR transgenic mice.

    Cells were seeded onto 6-well plates at 2×105 cells per well overnight. They were then transfected with 1 µg of VerUTR or control vector in combination with scrambled RNA or siRNA against VerUTR. Proteins were extracted 48 hours after transfection by lysing in 60 µl of lysis buffer containing protease inhibitors (150 mM NaCl, 25 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 1% Triton X-100, 8 M Urea, and 1x protease inhibitor cocktail). Tissues were disrupted in appropriate volume of lysis buffer depending on tissue weight. All samples were subjected to SDS-PAGE and then transferred to nitrocellulose membranes followed by incubating with ab24 at 1/500 dilution at 4°C overnight. The secondary antibody used was goat anti-mouse IgG at 1/2000 dilution at room temperatu

References for Anti-Rb antibody [Rb1 1F8] (ab24)

This product has been referenced in:
  • Araki K  et al. Cytoplasmic translocation of the retinoblastoma protein disrupts sarcomeric organization. Elife 2:e01228 (2013). IHC-Fr ; Human . Read more (PubMed: 24302570) »
  • Wan YW  et al. A smoking-associated 7-gene signature for lung cancer diagnosis and prognosis. Int J Oncol 41:1387-96 (2012). WB ; Human . Read more (PubMed: 22825454) »

See all 9 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Jun 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (lung and foreskin fibroblast cells; MOUSE lung)
Loading amount
30 µg
Specification
lung and foreskin fibroblast cells; MOUSE lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Apr 10 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Specification
Hela
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Mrs. Bernadette Ferraro

Verified customer

Submitted Dec 19 2005

Thank you for your enquiry. Ab24 is predicted to react with rat Rb (97% identity with immunogen) due to sequence homology but this has not been specifically tested yet. Should you decide to go ahead and purchase this product, please let us know how ...

Read More

Thank you for your enquiry. Ab24 also recognises the phosphorylates and unphospohorylated forms of Rb1, so it is nonspecific. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (human fibroblast)
Specification
human fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Oct 05 2005

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up