Overview

  • Product nameAnti-pan Arrestin antibody
  • Description
    Rabbit polyclonal to pan Arrestin
  • SpecificityAb2914 detects recombinant rat beta-arrestin and beta-arrestin2 (not tested against endogenous protein). This antibody does not detect visual or cone arrestin.
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Rat, Cow, Human
    Predicted to work with: Mouse, Rabbit, Cynomolgus Monkey, Rainbow Trout
  • Immunogen

    Synthetic peptide corresponding to Human pan Arrestin aa 384-397.
    Sequence:

    DDIVFEDFARLRLK


    (Peptide available as ab4932)

  • Positive control
    • COS7 cell lysates overexpressing rat beta arrestin 2 (flag tagged)

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Primary antibody notesVision involves the conversion of light into electrochemical signals that are processed by the retina and subsequently sent to and interpreted by the brain. The process of converting light to an electrochemical signal begins when the membrane-bound protein, rhodopsin, absorbs light within the retina. Photoexcitation of rhodopsin causes the cytoplasmic surface of the protein to become catalytically active. In the active state, rhodopsin activates transducin, a GTP binding protein. Once activated, transducin promotes the hydrolysis of cGMP by phosphodiesterase (PDE). The decrease of intracellular cGMP concentrations causes the ion channels within the outer segment of the rod or cone to close, thus causing membrane hyperpolarization and, eventually, signal transmission. Rhodopsin’s activity is believed to be shut off by its phosphorylation followed by binding of the soluble protein arrestin. Arrestins are cytosolic proteins that are involved in G protein-coupled receptor (GPCR) desensitization. Arrestin binding to activated GPCRs is phosphorylation dependent and, once bound, uncouple the GPCR from the associated heterotrimeric G proteins. There are currently 4 known mammalian isoforms, beta-arrestin1 (arrestin2), beta-arrestin2 (arrestin3), visual arrestin (arrestin1), and cone arrestin. The beta- isoforms are ubiquitously expressed and are known to interact with acetylcholine and adrenergic receptors. Visual and cone arrestins are found to interact directly with transducin.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab2914 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
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 3 µg/ml. Detects a band of approximately 47, 49 kDa.Can be blocked with Human pan Arrestin peptide (ab4932).
IHC-P 1/100 - 1/1000.
IP Use at an assay dependent concentration.

Target

  • FunctionFunctions in regulating agonist-mediated G-protein coupled receptor (GPCR) signaling by mediating both receptor desensitization and resensitization processes. During homologous desensitization, beta-arrestins bind to the GPRK-phosphorylated receptor and sterically preclude its coupling to the cognate G-protein; the binding appears to require additional receptor determinants exposed only in the active receptor conformation. The beta-arrestins target many receptors for internalization by acting as endocytic adapters (CLASPs, clathrin-associated sorting proteins) and recruiting the GPRCs to the adapter protein 2 complex 2 (AP-2) in clathrin-coated pits (CCPs). However, the extent of beta-arrestin involvement appears to vary significantly depending on the receptor, agonist and cell type. Internalized arrestin-receptor complexes traffic to intracellular endosomes, where they remain uncoupled from G-proteins. Two different modes of arrestin-mediated internalization occur. Class A receptors, like ADRB2, OPRM1, ENDRA, D1AR and ADRA1B dissociate from beta-arrestin at or near the plasma membrane and undergo rapid recycling. Class B receptors, like AVPR2, AGTR1, NTSR1, TRHR and TACR1 internalize as a complex with arrestin and traffic with it to endosomal vesicles, presumably as desensitized receptors, for extended periods of time. Receptor resensitization then requires that receptor-bound arrestin is removed so that the receptor can be dephosphorylated and returned to the plasma membrane. Involved in internalization of P2RY4 and UTP-stimulated internalization of P2RY2. Involved in phosphorylation-dependent internalization of OPRD1 ands subsequent recycling. Involved in the degradation of cAMP by recruiting cAMP phosphodiesterases to ligand-activated receptors. Beta-arrestins function as multivalent adapter proteins that can switch the GPCR from a G-protein signaling mode that transmits short-lived signals from the plasma membrane via small molecule second messengers and ion channels to a beta-arrestin signaling mode that transmits a distinct set of signals that are initiated as the receptor internalizes and transits the intracellular compartment. Acts as signaling scaffold for MAPK pathways such as MAPK1/3 (ERK1/2). ERK1/2 activated by the beta-arrestin scaffold is largely excluded from the nucleus and confined to cytoplasmic locations such as endocytic vesicles, also called beta-arrestin signalosomes. Recruits c-Src/SRC to ADRB2 resulting in ERK activation. GPCRs for which the beta-arrestin-mediated signaling relies on both ARRB1 and ARRB2 (codependent regulation) include ADRB2, F2RL1 and PTH1R. For some GPCRs the beta-arrestin-mediated signaling relies on either ARRB1 or ARRB2 and is inhibited by the other respective beta-arrestin form (reciprocal regulation). Inhibits ERK1/2 signaling in AGTR1- and AVPR2-mediated activation (reciprocal regulation). Is required for SP-stimulated endocytosis of NK1R and recruits c-Src/SRC to internalized NK1R resulting in ERK1/2 activation, which is required for the antiapoptotic effects of SP. Is involved in proteinase-activated F2RL1-mediated ERK activity. Acts as signaling scaffold for the AKT1 pathway. Is involved in alpha-thrombin-stimulated AKT1 signaling. Is involved in IGF1-stimulated AKT1 signaling leading to increased protection from apoptosis. Involved in activation of the p38 MAPK signaling pathway and in actin bundle formation. Involved in F2RL1-mediated cytoskeletal rearrangement and chemotaxis. Involved in AGTR1-mediated stress fiber formation by acting together with GNAQ to activate RHOA. Appears to function as signaling scaffold involved in regulation of MIP-1-beta-stimulated CCR5-dependent chemotaxis. Involved in attenuation of NF-kappa-B-dependent transcription in response to GPCR or cytokine stimulation by interacting with and stabilizing CHUK. May serve as nuclear messenger for GPCRs. Involved in OPRD1-stimulated transcriptional regulation by translocating to CDKN1B and FOS promoter regions and recruiting EP300 resulting in acetylation of histone H4. Involved in regulation of LEF1 transcriptional activity via interaction with DVL1 and/or DVL2 Also involved in regulation of receptors other than GPCRs. Involved in Toll-like receptor and IL-1 receptor signaling through the interaction with TRAF6 which prevents TRAF6 autoubiquitination and oligomerization required for activation of NF-kappa-B and JUN. Binds phosphoinositides. Binds inositolhexakisphosphate (InsP6) (By similarity). Involved in IL8-mediated granule release in neutrophils.
  • Sequence similaritiesBelongs to the arrestin family.
  • DomainThe [DE]-X(1,2)-F-X-X-[FL]-X-X-X-R motif mediates interaction the AP-2 complex subunit AP2B1 (By similarity). Binding to phosphorylated GPCRs induces a conformationanl change that exposes the motif to the surface.
    The N-terminus binds InsP6 with low affinity.
    The C-terminus binds InsP6 with high affinity.
  • Post-translational
    modifications
    Constitutively phosphorylated at Ser-412 in the cytoplasm. At the plasma membrane, is rapidly dephosphorylated, a process that is required for clathrin binding and ADRB2 endocytosis but not for ADRB2 binding and desensitization. Once internalized, is rephosphorylated.
    The ubiquitination status appears to regulate the formation and trafficking of beta-arrestin-GPCR complexes and signaling. Ubiquitination appears to occur GPCR-specific. Ubiquitinated by MDM2; the ubiquitination is required for rapid internalization of ADRB2. Deubiquitinated by USP33; the deubiquitination leads to a dissociation of the beta-arrestin-GPCR complex. Stimulation of a class A GPCR, such as ADRB2, induces transient ubiquitination and subsequently promotes association with USP33.
  • Cellular localizationCytoplasm. Nucleus. Cell membrane. Membrane > clathrin-coated pit. Cell projection > pseudopodium. Cytoplasmic vesicle. Translocates to the plasma membrane and colocalizes with antagonist-stimulated GPCRs. The monomeric form is predominantly located in the nucleus. The oligomeric form is located in the cytoplasm. Translocates to the nucleus upon stimulation of OPRD1.
  • Information by UniProt
  • Database links
  • Alternative names
    • ARB1 antibody
    • ARB2 antibody
    • ARR1 antibody
    • ARR2 antibody
    • ARRB1 antibody
    • ARRB1_HUMAN antibody
    • Arrestin beta 1 antibody
    • Arrestin beta 2 antibody
    • Arrestin beta-1 antibody
    • Beta-arrestin-1 antibody
    see all

Anti-pan Arrestin antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of pan Arrestin showing staining in the cytoplasm and nucleus of paraffin-treated human cerebellum tissue (right) compared with a negative control in the absence of primary antibody (left). Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then incubated with ab2914 diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of pan Arrestin showing staining in the cytoplasm and nucleus of paraffin-treated rat brain tissue (right) compared with a negative control in the absence of primary antibody (left). Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then incubated with ab2914 diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of pan Arrestin showing staining in the cytoplasm and nucleus of paraffin-treated rat spleen tissue (right) compared with a negative control in the absence of primary antibody (left). Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then incubated with ab2914 diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • All lanes : Anti-pan Arrestin antibody (ab2914) at 1/1000 dilution

    Lane 1 : C6 cell lysate
    Lane 2 : Rat brain cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 25 µg per lane.
  •  ab2914 staining human HEK 293 cells by ICC/IF. The cells were fixed in 4% PFA (in PBS) for 30min. After washing the cells were permeabilised with 0.01% Triton X-100 and then incubated with the primary antibody in PBS-T for 18h at 4C. An Alexa Fluor ® 488 conjugated donkey polyclonal antibody was used as the secondary (staining shown in green). After three 5min washes with PBS the cover slips were mounted with slow fade mounting solution containing DAPI (blue) .

    See Abreview

  • Anti-pan Arrestin antibody (ab2914) at 1 µg/ml + Human brain tissue lysate - total protein (ab29466) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 49 +51 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 27 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 90 seconds
    pan Arrestin contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

References for Anti-pan Arrestin antibody (ab2914)

This product has been referenced in:
  • Law IK  et al. Neurotensin-induced pro-inflammatory signaling in human colonocytes is regulated by beta-arrestins and endothelin-converting enzyme-dependent endocytosis and re-sensitization of NT receptor 1. J Biol Chem : (2012). Read more (PubMed: 22416137) »
  • Kuhr FK  et al. Beta-arrestin 2 is required for B1 receptor-dependent post-translational activation of inducible nitric oxide synthase. FASEB J 24:2475-83 (2010). WB ; Human . Read more (PubMed: 20228252) »

See all 7 Publications for this product

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Thank you for your enquiry.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEK 293)
Specification HEK 293
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
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Mrs. Danijela Markovic

Verified customer

Submitted Aug 30 2006

Thank you very much for your protocol details, I think there may be a step which will also help you (as well as running the positive controls mentioned in my previous e-mail-brain and spleen). In our experience multiple non specific bands can be cause...

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I have been informed this morning that the antibody was tested in WB against COS7 cell lysates overexpressing rat beta arrestin 2 (flag tagged) and have not therefore been tested against the endogenous protein. A literature search revealed that in huma...

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I'm sorry to hear you are experiencing problem with ab2914. We have not received other complaints since 2004 about this antibody and have not tested it in human samples but having done a BLAST search I found 100% homology of the immunogen with the huma...

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I am sorry to hear that you are experiencing trouble with this antibody. I have enquired with the originator of ab2914 and everything appears fine with your protocol. We have had no other complaints about this antibody, but this particular antibody has...

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