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Signal Transduction Signaling Pathway G Protein Signaling Heterotrimeric G Proteins Regulators
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Anti-Beta Arrestin 2 antibody (ab31294)

  • Datasheet
  • SDS
Reviews (7)Q&A (4)References (16)

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Western blot - Anti-Beta Arrestin 2 antibody (ab31294)
  • Flow Cytometry - Anti-Beta Arrestin 2 antibody (ab31294)
  • Immunocytochemistry - Anti-Beta Arrestin 2 antibody (ab31294)

Key features and details

  • Goat polyclonal to Beta Arrestin 2
  • Suitable for: WB, Flow Cyt, ICC
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Beta Arrestin 2 antibody [EPR22073] (ab206972)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Beta Arrestin 2 antibody
    See all Beta Arrestin 2 primary antibodies
  • Description

    Goat polyclonal to Beta Arrestin 2
  • Host species

    Goat
  • Specificity

    This antibody is expected to recognise both reported isoforms (NP_004304 and NP_945355). No cross reactivity is expected with Arrestin beta 1.
  • Tested applications

    Suitable for: WB, Flow Cyt, ICCmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide:

    C-HDHIPLPRPQS

    , corresponding to Internal sequence amino acids 350-360 of Human Beta Arrestin 2.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • ICC: HeLa cells; Flow Cyt: A549 cells WB: A549, U251 cells
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    This antibody was purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • Heterotrimeric G Proteins
    • Regulators

Associated products

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant Human Beta Arrestin 2 protein (ab132309)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab31294 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 (5)
Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 40-48 kDa (predicted molecular weight: 46 kDa).

A 1 hour primary incubation is recommended for this product.

Flow Cyt
Use a concentration of 10 µg/ml.
ICC
Use a concentration of 10 µg/ml.
Notes
WB
Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 40-48 kDa (predicted molecular weight: 46 kDa).

A 1 hour primary incubation is recommended for this product.

Flow Cyt
Use a concentration of 10 µg/ml.
ICC
Use a concentration of 10 µg/ml.

Target

  • Function

    Functions 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. Mediates endocytosis of CCR7 following ligation of CCL19 but not CCL21. Involved in internalization of P2RY1, P2RY4, P2RY6 and P2RY11 and ATP-stimulated internalization of P2RY2. Involved in phopshorylation-dependent internalization of OPRD1 and subsequent recycling or degradation. Involved in ubiquitination of IGF1R. 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) and MAPK10 (JNK3). ERK1/2 and JNK3 activated by the beta-arrestin scaffold are largely excluded from the nucleus and confined to cytoplasmic locations such as endocytic vesicles, also called beta-arrestin signalosomes. Acts as signaling scaffold for the AKT1 pathway. 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). Increases ERK1/2 signaling in AGTR1- and AVPR2-mediated activation (reciprocal regulation). Involved in CCR7-mediated ERK1/2 signaling involving ligand CCL19. Is involved in type-1A angiotensin II receptor/AGTR1-mediated ERK activity. Is involved in type-1A angiotensin II receptor/AGTR1-mediated MAPK10 activity. Is involved in dopamine-stimulated AKT1 activity in the striatum by disrupting the association of AKT1 with its negative regulator PP2A. Involved in AGTR1-mediated chemotaxis. 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. Suppresses UV-induced NF-kappa-B-dependent activation by interacting with CHUK. The function is promoted by stimulation of ADRB2 and dephosphorylation of ARRB2. Involved in p53/TP53-mediated apoptosis by regulating MDM2 and reducing the MDM2-mediated degradation of p53/TP53. May serve as nuclear messenger for GPCRs. Upon stimulation of OR1D2, may be involved in regulation of gene expression during the early processes of fertilization. Also involved in regulation of receptors others than GPCRs. Involved in endocytosis of TGFBR2 and TGFBR3 and down-regulates TGF-beta signaling such as NF-kappa-B activation. Involved in endocytosis of low-density lipoprotein receptor/LDLR. Involved in endocytosis of smoothened homolog/Smo, which also requires ADRBK1. Involved in endocytosis of SLC9A5. Involved in endocytosis of ENG and subsequent TGF-beta-mediated ERK activation and migration of epithelial cells. 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. Involved in insulin resistence by acting as insulin-induced signaling scaffold for SRC, AKT1 and INSR. Involved in regulation of inhibitory signaling of natural killer cells by recruiting PTPN6 and PTPN11 to KIR2DL1.
  • Sequence similarities

    Belongs to the arrestin family.
  • Domain

    The [DE]-X(1,2)-F-X-X-[FL]-X-X-X-R motif mediates interaction the AP-2 complex subunit AP2B1.
  • Post-translational
    modifications

    Phosphorylated at Thr-382 in the cytoplasm; probably dephosphorylated at the plasma membrane. The phosphorylation does not regulate internalization and recycling of ADRB2, interaction with clathrin or AP2B1.
    The ubiquitination status appears to regulate the formation and trafficking of beta-arrestin-GPCR complexes and signaling. Ubiquitination appears to occurr GPCR-specifc. 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. Stimulation of a class B GPCR promotes a sustained ubiquitination.
  • Cellular localization

    Cytoplasm. Nucleus. Cell membrane. Membrane > clathrin-coated pit. Cytoplasmic vesicle. Translocates to the plasma membrane and colocalizes with antagonist-stimulated GPCRs.
  • Target information above from: UniProt accession P32121 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 409 Human
    • Omim: 107941 Human
    • SwissProt: P32121 Human
    • Unigene: 435811 Human
    • Alternative names

      • ARB 2 antibody
      • ARB2 antibody
      • ARR 2 antibody
      • ARR2 antibody
      • ARRB 2 antibody
      • ARRB2 antibody
      • ARRB2_HUMAN antibody
      • Arrestin 3 antibody
      • Arrestin beta 2 antibody
      • Arrestin beta-2 antibody
      • BARR2 antibody
      • Beta-arrestin-2 antibody
      • DKFZp686L0365 antibody
      • HGNC:712 antibody
      see all

    Images

    • Western blot - Anti-Beta Arrestin 2 antibody (ab31294)
      Western blot - Anti-Beta Arrestin 2 antibody (ab31294)
      All lanes : Anti-Beta Arrestin 2 antibody (ab31294) at 1 µg/ml

      Lane 1 : A549 (human lung carcinoma cell line) cell lysate
      Lane 2 : U251 (human brain glioma cell line) cell lysate

      Predicted band size: 46 kDa

    • Flow Cytometry - Anti-Beta Arrestin 2 antibody (ab31294)
      Flow Cytometry - Anti-Beta Arrestin 2 antibody (ab31294)

      Flow cytometric analysis of paraformaldehyde-fixed 0.5% Triton-permeabilized A549 cells (human lung carcinoma cell line) staining beta arrestin positive cells with 10 µg/ml ab31294 and 1 µg/ml Alexa Fluor 488 secondary antibody (blue) and negative control unimmunized goat IgG (black). 

    • Immunocytochemistry - Anti-Beta Arrestin 2 antibody (ab31294)
      Immunocytochemistry - Anti-Beta Arrestin 2 antibody (ab31294)

      Immunocytochemistry immunofluorescence analysis of paraformaldehyde-fixed 0.15% Triton-permeabilized HeLa cells staining beta arrestin 2 with 10 µg/ml ab31294 and Alexa Fluor 488 secondary antibody 20 µg/ml; plasma membrane and cytoplasmic staining is in green, and the nucleus is counterstained with DAPI in blue. The negative control is unimmunized goat IgG.

    Protocols

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (16)

    Publishing research using ab31294? Please let us know so that we can cite the reference in this datasheet.

    ab31294 has been referenced in 16 publications.

    • Philip JL  et al. Regulation of cardiac fibroblast-mediated maladaptive ventricular remodeling by ß-arrestins. PLoS One 14:e0219011 (2019). PubMed: 31269046
    • Zhang XR  et al. ß-arrestin2 alleviates L-dopa-induced dyskinesia via lower D1R activity in Parkinson's rats. Aging (Albany NY) 11:12315-12327 (2019). PubMed: 31891566
    • Zhang ZR  et al. Striatal overexpression of ß-arrestin2 counteracts L-dopa-induced dyskinesia in 6-hydroxydopamine lesioned Parkinson's disease rats. Neurochem Int 131:104543 (2019). PubMed: 31491493
    • Eng AG  et al. Transduction of group I mGluR-mediated synaptic plasticity by ß-arrestin2 signalling. Nat Commun 7:13571 (2016). IP ; Mouse . PubMed: 27886171
    • Ying L  et al. The transient receptor potential vanilloid 4 channel modulates uterine tone during pregnancy. Sci Transl Med 7:319ra204 (2015). PubMed: 26702092
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 11 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Beta Arrestin 2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK293)
    Permeabilization
    Yes - 0.1% Triton X100
    Specification
    HEK293
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 30 2017

    Immunoprecipitation abreview for Anti-Beta Arrestin 2 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Other - Pierce Protein A/G Magnetic Beads
    Sample
    Mouse Cell lysate - whole cell (Heart)
    Specification
    Heart
    Total protein in input
    50 µg
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 09 2014

    Western blot abreview for Anti-Beta Arrestin 2 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Mouse Tissue lysate - whole (Mouse brain)
    Specification
    Mouse brain
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Victor Maximov

    Verified customer

    Submitted May 26 2014

    Question

    I will let you know what is coming out. Have a great weekend,

    Read More

    Abcam community

    Verified customer

    Asked on Jun 22 2012

    Answer

    You are very welcome.

    I look forward to hearing from you.

    Have a great weekend as well.

    Read More

    Abcam Scientific Support

    Answered on Jun 22 2012

    Question

    We ordered the anti- beta-arrestin 1 and anti- beta-arrestin 2 antibodies from your company recently. The information is as below: Rabbit monoclonal[E274] to beta-arrestin 1 (ab32099) Lot# GR37458-5 Goat polyclonal (ab31294) for the beta-arrestin 2 Lot#GR77572-1 The beta-arrestin1 antibodies works by showing that knockdown of beta-arrestin1 decrease the proper size band. But the anti-beta-arrestin2 is not working (no bands showed up even after overexpression of beta-arrestin2 protein in the cells). I am wondering if you might kindly send us some positive control for beta-arrestin 2 or a new batch of antibody? Thanks & I am looking forward to your reply!

    Read More

    Abcam community

    Verified customer

    Asked on Jun 21 2012

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry to hear about the difficulties which you have exprienced when using this product and for any inconvenience this has caused. As requested, I have issued a free of charge positive control lysate, ab29466 Brain (normal) Tissue Lysate with the order number xxxxx. To check the status of the order please contact our Customer Service team and reference this number. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Jun 21 2012

    Question

    I used western blot to test the heart fibroblast beta-arrestins expression. The normal RIPA buffer was used to lysis the cells (which was also added with PMSF, Protein kinase inhibitor cocktail). After blotting 30 ug of the protein onto PVDF membrane, the membrane was developed for the western. (5% BSA was used as a blocking buffer and antibody dilution buffer; 1st antibody was developing at 4 degree overnight (1:500 dilution), and 2nd antibody was developing at RT for 2 hours). But finally I could not get any bands for the beta-arrestin2 antibody in the proper size site. Could you kindly send us the positive control to help us to find out if the antibody is working or not. Or we need change to another batch or different Cat# antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 21 2012

    Answer

    Thank you for your reply and for these additional details.

    Would you be able to send me the Abcam order confirmation number for this purchase? I will need this information for replacement, refund or for sending a positive control to you.

    I have one follow up question regarding this issue. You say thatcould not get any bands for the beta-arrestin 2 antibody in the proper size site. Are you seeing other bands? If so where are these bands located? Would you be able to send an image of your blot to me?

    This product is covered by our Abpromise guarantee. If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.

    I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

    Use our products? Submit an Abreview. Earn rewards!

    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Jun 21 2012

    Question

    Dear Sir/Madam, We ordered the anti- beta-arrestin 1 and anti- beta-arrestin 2 antibodies from your company recently. The information is as below: Rabbit monoclonal[E274] to beta-arrestin 1 (ab32099) Lot# GR37458-5 Goat polyclonal (ab31294) for the beta-arrestin 2 Lot#GR77572-1 The beta-arrestin1 antibodies works by showing that knockdown of beta-arrestin1 decrease the proper size band. But the anti-beta-arrestin2 is not working (no bands showed up even after overexpression of beta-arrestin2 protein in the cells). I am wondering if you might kindly send us some positive control for beta-arrestin 2 or a new batch of antibody? Thanks & I am looking forward to your reply!

    Read More

    Abcam community

    Verified customer

    Asked on Jun 20 2012

    Answer

    Thank you for contacting Abcam.

    I am sorry to hear of the difficulties that you have been experiencing using these products and would like to help you to remedy this. Would you mind sending me the details of your experiments such as species type, sample preparation and protocol including any steps that you may have changed while troubleshooting?

    These details will not only help us to better understand the difficulties you are experiencing but will also be used for internal testing of the products if deemed necessary.



    We guarantee all our products for TESTED species and applications as listed on the datasheet. The guarantee is valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: https://www.abcam.com/abpromise.

    Please let me know if you have any questions.

    Read More

    Abcam Scientific Support

    Answered on Jun 21 2012

    Western blot abreview for Anti-Beta Arrestin 2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Colorectal cancer cell line SW480)
    Loading amount
    20 µg
    Specification
    Colorectal cancer cell line SW480
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 02 2008

    Western blot abreview for Anti-Beta Arrestin 2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (aortic smooth muscle cell)
    Loading amount
    15 µg
    Specification
    aortic smooth muscle cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 04 2007

    Western blot abreview for Anti-Beta Arrestin 2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (smooth muscle cells)
    Loading amount
    15 µg
    Specification
    smooth muscle cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 23 2007

    1-10 of 11 Abreviews or Q&A

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