Overview

  • Product name

    Rap1 Activation Assay Kit
  • Sample type

    Tissue, Adherent cells, Suspension cells
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    Rap1 Activation Assay Kit (ab212011) uses RalGDS RBD Agarose beads to selectively isolate and pull-down the active form of Rap from purified samples or endogenous lysates.  Subsequently, the precipitated GTP-Rap is detected by western blot analysis using an anti-Rap1 polyclonal antibody, which reacts with the human, mouse and rat Rap1A and Rap1B proteins.


    Features: 1) non radioactive assay format; 2) fast results: 1 hour assay plus electrophoresis/blotting time; 3) includes Cdc42 positive control; 4) pink colored agarose beads for easy identification during washing and aspiration steps.

  • Notes

    Small GTP-binding proteins (or GTPases) are a family of proteins that serve as molecular regulators in signaling transduction pathways.

    Rap1 (Ras-related protein 1), a 21 kDa protein of the Ras superfamily, exhibits 60% identity to Rap2. Rap2 regulates a variety of biological response pathways that include cell migration, cell adhesion, and embryonic blood vessel formation. Like other small GTPases, Rap1 regulates molecular events by cycling between an inactive GDP-bound form and an active GTP-bound form. In its active (GTP-bound) state, Rap1 bind specifically to the Rap-binding domain (RBD) of RalGDS to control downstream signaling cascades.

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 20 tests 5 tests
    ab211179 - RalGDS RBD Agarose Beads 1 x 800µl 1 x 200µl
    100X GTPyS 1 x 50µl 1 x 20µl
    100X GDP 1 x 50µl 1 x 20µl
    5X Assay/Lysis Buffer 1 x 30ml 4 x 2ml
    Anti-Rap1 Goat polyclonal antibody 1 x 40µl 1 x 20µl
    GTPase immunoblot Positive Control 1 x 100µl 1 x 100µl
  • Research areas

  • Function

    Acts both as a regulator of telomere function and as a transcription regulator. Involved in the regulation of telomere length and protection as a component of the shelterin complex (telosome). In contrast to other components of the shelterin complex, it is dispensible for telomere capping and does not participate in the protection of telomeres against non-homologous end-joining (NHEJ)-mediated repair. Instead, it is required to negatively regulate telomere recombination and is essential for repressing homology-directed repair (HDR), which can affect telomere length. Does not bind DNA directly: recruited to telomeric double-stranded 5'-TTAGGG-3' repeats via its interaction with TERF2. Independently of its function in telomeres, also acts as a transcription regulator: recruited to extratelomeric 5'-TTAGGG-3' sites via its association with TERF2 or other factors, and regulates gene expression. When cytoplasmic, associates with the I-kappa-B-kinase (IKK) complex and acts as a regulator of the NF-kappa-B signaling by promoting IKK-mediated phosphorylation of RELA/p65, leading to activate expression of NF-kappa-B target genes.
  • Tissue specificity

    Ubiquitous. Highly expressed.
  • Sequence similarities

    Belongs to the RAP1 family.
    Contains 1 BRCT domain.
    Contains 1 Myb-like domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus. Cytoplasm. Chromosome. Chromosome > telomere. Associates with chromosomes, both at telomeres and in extratelomeric sites. Also exists as a cytoplasmic form, where it associates with the IKK complex.
  • Information by UniProt
  • Alternative names

    • Dopamine receptor interacting protein 5
    • Dopamine receptor-interacting protein 5
    • DRIP 5
    • DRIP5
    • hRap1
    • MGC105533
    • RAP 1
    • RAP1 homolog
    • RAP1, yeast, homolog if
    • RAP1, yeast, homolog of
    • Repressor/activator protein 1 homolog
    • TE2IP_HUMAN
    • Telomeric Repeat Binding Factor 2 Interacting Protein
    • Telomeric repeat-binding factor 2-interacting protein 1
    • TERF2-interacting protein
    • TERF2-interacting telomeric protein 1
    • TERF2IP
    • TRF2 Interacting Telomeric Protein RAP1
    • TRF2 interacting telomeric RAP1 protein
    • TRF2-interacting telomeric protein
    • TRF2-interacting telomeric protein 1
    see all
  • Database links

Associated products

Images

  • Rap1 positive control (10 µL of NIH3T3 cell lysate at 0.75 mg/mL) detected by immunoblot.

  • Rap1 activation assay. Lane 1: NIH 3T3 cell lysate loaded with GDP and incubated with RalGDS RBD Agarose beads. Lane 2: NIH 3T3 cell lysate loaded with GTPγS and incubated with RalGDS RBD Agarose beads.

Protocols

References

ab212011 has not yet been referenced specifically in any publications.

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