Recombinant Human Rb protein (ab83205)

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Western blot - Recombinant Human Rb protein (ab83205)

    Key features and details

    • Expression system: Baculovirus
    • Purity: > 95% SDS-PAGE
    • Tags: His tag N-Terminus
    • Suitable for: SDS-PAGE, GSA, WB

    Description

    • Product name

      Recombinant Human Rb protein
      See all Rb proteins and peptides

    • Purity

      > 95 % SDS-PAGE.
      Purified by affinity and FPLC chromatography.

    • Expression system

      Baculovirus

    • Protein length

      Full length protein

    • Animal free

      No

    • Nature

      Recombinant

      • Species

        Human

      • Sequence

        TPRKTAATAA AAAAEPPAPP PPPPPEEDPE QDSGPEDLPL VRLEFEETEE PDFTALCQKL KIPDHVRERA WLTWEKVSSV DGVLGGYIQK KKELWGICIF IAAVDLDEMS FTFTELQKNI EISVHKFFNL LKEIDTSTKV DNAMSRLLKK YDVLFALFSK LERTCELIYL TQPSSSISTE INSALVLKVS WITFLLAKGE VLQMEDDLVI SFQLMLCVLD YFIKLSPPML LKEPYKTAVI PINGSPRTPR RGQNRSARIA KQLENDTRII EVLCKEHECN IDEVKNVYFK NFIPFMNSLG LVTSNGLPEV ENLSKRYEEI YLKNKDLDAR LFLDHDKTLQ TDSIDSFETQ RTPRKSNLDE EVNVIPPHTP VRTVMNTIQQ LMMILNSASD QPSENLISYF NNCTVNPKES ILKRVKDIGY IFKEKFAKAV GQGCVEIGSQ RYKLGVRLYY RVMESMLKSE EERLSIQNFS KLLNDNIFHM SLLACALEVV MATYSRSTSQ NLDSGTDLSF PWILNVLNLK AFDFYKVIES FIKAEGNLTR EMIKHLERCE HRIMESLAWL SDSPLFDLIK QSKDREGPTD HLESACPLNL PLQNNHTAAD MYLSPVRSPK KKGSTTRVNS TANAETQATS AFQTQKPLKS TSLSLFYKKV YRLAYLRLNT LCERLLSEHP ELEHIIWTLF QHTLQNEYEL MRDRHLDQIM MCSMYGICKV KNIDLKFKII VTAYKDLPHA VQETFKRVLI KEEEYDSIIV FYNSVFMQRL KTNILQYAST RPPTLSPIPH IPRSPYKFPS SPLRIPGGNI YISPLKSPYK ISEGLPTPTK MTPRSRILVS IGESFGTSEK FQKINQMVCN SDRVLKRSAE GSNPPKPLKK LRFDIEGSDE ADGSKHLPGE SKFQQKLAEM TSTRTRMQKQ KMNDSMDTSN KEEK

      • Predicted molecular weight

        108 kDa

      • Amino acids

        1 to 928

      • Tags

        His tag N-Terminus

    Specifications

    Our Abpromise guarantee covers the use of ab83205 in the following tested applications.

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

    • Applications

      SDS-PAGE

      Gel Supershift Assays

      Western blot

      EMSA

    • Form

      Liquid
    • Concentration information loading...

    Preparation and Storage

    • Stability and Storage

      Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

      pH: 7.9
      Constituents: 0.0154% DTT, 0.316% Tris HCl, 0.00584% EDTA, 20% Glycerol (glycerin, glycerine)

    General Info

    • Alternative names

      • Exon 17 tumor GOS561 substitution mutation causes premature stop
      • GOS563 exon 17 substitution mutation causes premature stop
      • OSRC
      • Osteosarcoma
      • p105-Rb
      • P105RB
      • PP105
      • pp110
      • PPP1R130
      • pRb
      • Prepro retinoblastoma associated protein
      • Protein phosphatase 1 regulatory subunit 130
      • Rb
      • RB transcriptional corepressor 1
      • RB_HUMAN
      • RB1
      • RB1 gene
      • Retinoblastoma 1
      • Retinoblastoma suspectibility protein
      • Retinoblastoma-associated protein
      see all

    • 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

    Images

    • Western blot - Recombinant Human Rb protein (ab83205)
      Western blot - Recombinant Human Rb protein (ab83205)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab83205? Please let us know so that we can cite the reference in this datasheet.

    ab83205 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    Re: Reply from Abcam to your enquiry regarding ab83205 [CCE3194263] Thanks! If I could get a copy of the image it would be great. I used the purified Rb from abcam for a DNAse I protection assay. Rb doesn’t associate to DNA on its own, it requires interaction with proteins like E2Fs or SP1. So with my DNAse I protection assay I expected to see no protection when incubated with purified Rb (ie Rb without any other protein contanminants). Surprisingly I saw a precise protection pattern similar to that I observed upon incubation with SP1 which puzzled me. So that’s when I emailed abcam to determine if there were any other proteins with the purified Rb. Rb’s interaction with E2Fs and SP1 is very strong therefore based on my results I thought that there may be some contamination of either of these proteins in the purified Rb mixture. Hope this helps let me know if you need anymore info. Thanks

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    Answer

    Thanks for your reply. I have arranged for the lab to send a free of charge vial from the new lot, which should arrive in the next few days. Please see attached for the SDS-PAGE and Western blot images of the new lot of Rb protein. In regards to where the contaminating bands were seen in the previous lots, I received this reply: "Regarding the extra bands' MW, 3 weak bands are seen between 66 and 97 kDa markers and one in between 45 and 66 kDa". The lab has also asked if you could send any data from your transcription assay for their analysis. Please let me know if you have any questions, and I will update you shortly with an expected arrival data for the new vial of ab83205.

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    Question

    Hi, Thanks for your reply that explains a lot. Could you please tell me the sizes of the bands you see? I would truly appreciate if you could send a vial of the new lot with which I can repeat my experiments. Thanks

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    Answer

    Thank you for your reply. I will ask the lab for details about the bands that they saw. I have not seen an image myself. For our records, can you tell me what kind of results you obtained with this protein in your transcription assay? I will have a new vial sent to you as soon as possible. I look forward to hearing from you.

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    Question

    Hi I would like to know if there are any contaminating proteins in ab83205?

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    Answer

    Thank you for your phone call last month and for your patience while the lab has tested this protein. The lab tested the current lots of the protein and found that there were 3-4 light contaminant bands below the pRB (apparent Mw 107 kDa). The identities of these bands are unknown. The lab has just produced a new lot and removed most of the contaminating bands. If you are still interested, please let me know and I will get a vial of this new lot for you. Let me know if you have any questions, or if there is anything else that we can do for you.

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