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    aurora-b-antibody-ab2254.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Aurora
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Anti-Aurora B antibody (ab2254)

  • Datasheet
  • SDS
Reviews (36)Q&A (22)References (154)

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Western blot - Anti-Aurora B antibody (ab2254)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
  • Western blot - Anti-Aurora B antibody (ab2254)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody (ab2254)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody (ab2254)
  • Western blot - Anti-Aurora B antibody (ab2254)
  • Western blot - Anti-Aurora B antibody (ab2254)

Key features and details

  • Rabbit polyclonal to Aurora B
  • Suitable for: ICC/IF, IHC-P, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Aurora B antibody
    See all Aurora B primary antibodies
  • Description

    Rabbit polyclonal to Aurora B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Hamster, Pig
  • Immunogen

    Synthetic peptide within Human Aurora B aa 1-100 conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab13569)

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped 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 buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Aurora
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Phosphorylation
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Aurora
    • Cancer
    • Cell cycle
    • Kinases/phosphatases
    • Aurora

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab6721)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Aurora B protein (ab51435)

Applications

Our Abpromise guarantee covers the use of ab2254 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 a concentration of 0.5 - 1 µg/ml.

Methanol fixation recommended.

IHC-P 1/200.
WB 1/1000 - 1/2000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).

Target

  • Function

    May be directly involved in regulating the cleavage of polar spindle microtubules and is a key regulator for the onset of cytokinesis during mitosis. Component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly. Phosphorylates 'Ser-10' and 'Ser-28' of histone H3 during mitosis. Required for kinetochore localization of BUB1 and SGOL1. Interacts with INCENP.
  • Tissue specificity

    High level expression seen in the thymus. It is also expressed in the spleen, lung, testis, colon, placenta and fetal liver. Expressed during S and G2/M phase and expression is up-regulated in cancer cells during M phase.
  • Involvement in disease

    Note=Disruptive regulation of expression is a possibile mechanism of the perturbation of chromosomal integrity in cancer cells through its dominant-negative effect on cytokinesis.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Ubiquitinated by different BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complexes. Ubiquitinated by the BCR(KLHL9-KLHL13) E3 ubiquitin ligase complex, ubiquitination leads to removal from mitotic chromosomes and is required for cytokinesis. During anaphase, the BCR(KLHL21) E3 ubiquitin ligase complex recruits the CPC complex from chromosomes to the spindle midzone and mediates the ubiquitination of AURKB. Ubiquitination of AURKB by BCR(KLHL21) E3 ubiquitin ligase complex may not lead to its degradation by the proteasome.
  • Cellular localization

    Nucleus. Chromosome. Chromosome > centromere. Cytoplasm > cytoskeleton > spindle. Localizes on chromosome arms and inner centromeres from prophase through metaphase and then transferring to the spindle midzone and midbody from anaphase through cytokinesis. Colocalized with gamma tubulin in the mid-body.
  • Target information above from: UniProt accession Q96GD4 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 9212 Human
    • Entrez Gene: 20877 Mouse
    • Entrez Gene: 396955 Pig
    • Entrez Gene: 114592 Rat
    • Omim: 604970 Human
    • SwissProt: Q96GD4 Human
    • SwissProt: O70126 Mouse
    • SwissProt: Q9N0X0 Pig
    • SwissProt: O55099 Rat
    • Unigene: 442658 Human
    • Unigene: 3488 Mouse
    • Unigene: 10865 Rat
    see all
  • Alternative names

    • AIK2 antibody
    • AIM-1 antibody
    • AIM1 antibody
    • ARK-2 antibody
    • ARK2 antibody
    • AurB antibody
    • AURKB antibody
    • AURKB_HUMAN antibody
    • Aurora 1 antibody
    • Aurora and Ipl1 like midbody associated protein 1 antibody
    • Aurora kinase B antibody
    • Aurora related kinase 2 antibody
    • Aurora- and Ipl1-like midbody-associated protein 1 antibody
    • Aurora-B antibody
    • Aurora-related kinase 2 antibody
    • Aurora/IPL1 related kinase 2 antibody
    • Aurora/IPL1-related kinase 2 antibody
    • IPL1 antibody
    • PPP1R48 antibody
    • Protein phosphatase 1 regulatory subunit 48 antibody
    • Serine/theronine kinase 12 antibody
    • Serine/threonine protein kinase 12 antibody
    • Serine/threonine-protein kinase 12 antibody
    • Serine/threonine-protein kinase aurora-B antibody
    • STK-1 antibody
    • STK1 antibody
    • STK12 antibody
    • STK5 antibody
    see all

Images

  • Western blot - Anti-Aurora B antibody (ab2254)
    Western blot - Anti-Aurora B antibody (ab2254)Balboula and Schindler PLoS Genet. 2014 Feb 27;10(2):e1004194. doi: 10.1371/journal.pgen.1004194. eCollection 2014 Feb. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    AURKB is expressed in mouse oocytes.

    (Panel D) 20 GV-intact oocytes were collected from CF1 mice and micro-injected with the indicated cRNA. Two hours after injection, the oocytes were matured to Met II in vitro (16 h). The total numbers of non-injected control oocytes (Non-inj.) are indicated in parenthesis. Total cellular lysates were probed with the indicated antibody. The panels are images of the same membrane that was stripped and re-probed. The arrows indicate the specific AURKB protein band, and the asterisk indicates a presumed degradation product of AURKB-GFP.

  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)Balboula and Schindler PLoS Genet. 2014 Feb 27;10(2):e1004194. doi: 10.1371/journal.pgen.1004194. eCollection 2014 Feb. Fig 1.

    AURKB is expressed in mouse oocytes.

    (Panel A) GV-intact oocytes were collected from CF1 mice and matured in vitro for 8 h (Met I), or 16 h (Met II), prior to fixation and staining with an anti-AURKB antibody (ab2254).

  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)Chopra et al PLoS One. 2016 Apr 20;11(4):e0153818. doi: 10.1371/journal.pone.0153818. eCollection 2016. Fig 6. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    HCT 116 (Human colorectal carcinoma cell line) cells were examined by immunofluorescence confocal microscopy.

    Cells were treated with 500 nM AK301 for 16 hours, and then processed for Aurora B (ab2254) and β-tubulin staining (Panel B). The color key and 20 μm bars are shown. Images of representative field is shown with a 20 μm bar. End-labeled DNA is shown in red and DAPI-stained DNA is blue.

    Cells cultured on coverslips were fixed with 4% paraformaldehyde at room temperature or 100% ice cold methanol at 4°C and then permeabilized with 0.5% Triton X-100 in PBS. Cells were blocked in 5% serum (in PBS) and then incubated with primary antibody (in 5% serum) on shaker for 1 h at room temperature.

  • Western blot - Anti-Aurora B antibody (ab2254)
    Western blot - Anti-Aurora B antibody (ab2254)
    All lanes : Anti-Aurora B antibody (ab2254) at 1 µg/ml

    Lane 1 : HeLa cell lysate
    Lane 2 : HeLa nocodozole treated cell lysate
    Lane 3 : NIH3T3 cell lysate
    Lane 4 : PC12 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Performed under reducing conditions.

    Predicted band size: 39 kDa



    Blocked with 2% BSA.

  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)

    Immunofluorescence in human cells using Rabbit polyclonal to Aurora B (red), DAPI (blue) and CREST serum (binds to centromeres)(green).

    (a) HeLa cells - transition from interphase (left) through mitosis
    (b) RPE-1 cells - as in (a)
    (c) HeLa cells - interphase
    (d) RPE-1 cells - interphase

  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)

    ab2254 stained in Hela cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab2254 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody (ab2254)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody (ab2254)

    IHC image of Aurora B staining in Human Lymph node Hodgkins disease formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2254, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)
    Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody (ab2254)This image is courtesy of an Abreview from Lux Fatimathas.

    ab2254 staining human A431 (epithelial) cells by ICC/IF.  The sample was fixed in paraformaldehyde and permeabilized by incubation with 0.1% Triton X100.  1% BSA was used as the blocking agent prior to a 1 hour incubation with the primary antibody, diluted 1/1000 with 1% BSA made up in PBS.  An Alexa Fluor® 647 conjugated Donkey anti-Rabbit IgG (H+L) antibody was used as the secondary.  Blocking and antibody incubation steps were carried out at room temperature.

    In this set of images, the tubulin is stained green, Aurora B in pink and DNA in blue.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody (ab2254)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody (ab2254)

    Rabbit polyclonal to Aurora B (ab2254) used to stain SW620 human tumour xenografts (in mouse).

    The sections were microwave pretreated in citrate buffer (pH 6.0) for 5 mins high then 5 mins simmer (800W conventional microwave).  Slides were then incubated for 1 hour with the Aurora B primary antibody diluted 1/200 in TBS, then visualised using DAB, after application of an appropriate secondary. 

  • Western blot - Anti-Aurora B antibody (ab2254)
    Western blot - Anti-Aurora B antibody (ab2254)
    All lanes : Anti-Aurora B antibody (ab2254) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : HeLa Nuclear Lysate
    Lane 3 : Jurkat Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 39 kDa
    Observed band size: 39 kDa
    Additional bands at: 37 kDa (possible isoform)


    Exposure time: 150 seconds
  • Western blot - Anti-Aurora B antibody (ab2254)
    Western blot - Anti-Aurora B antibody (ab2254)
    Anti-Aurora B antibody (ab2254) at 1/2000 dilution + Recombinant human Aurora B protein (ab51435) at 0.1 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 39 kDa


    Exposure time: 30 seconds

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (154)

    Publishing research using ab2254? Please let us know so that we can cite the reference in this datasheet.

    ab2254 has been referenced in 154 publications.

    • Bywater MJ  et al. Reactivation of Myc transcription in the mouse heart unlocks its proliferative capacity. Nat Commun 11:1827 (2020). PubMed: 32286286
    • Salmina K  et al. "Mitotic Slippage" and Extranuclear DNA in Cancer Chemoresistance: A Focus on Telomeres. Int J Mol Sci 21:N/A (2020). PubMed: 32316332
    • Wang Y  et al. Single-cell analysis of murine fibroblasts identifies neonatal to adult switching that regulates cardiomyocyte maturation. Nat Commun 11:2585 (2020). PubMed: 32444791
    • Moteleub HNAE  et al. Predictors for hepatocellular carcinoma recurrence after microwave ablation. Clin Exp Hepatol 6:77-84 (2020). PubMed: 32728623
    • Su T  et al. Platelet-Inspired Nanocells for Targeted Heart Repair After Ischemia/Reperfusion Injury. Adv Funct Mater 29:N/A (2019). PubMed: 32256277
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-10 of 58 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-Aurora B antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Embryonic Heart)
    Permeabilization
    Yes - 0.2% Tween-20 for one hour
    Specification
    Embryonic Heart
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 16 2020

    Western blot abreview for Anti-Aurora B antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Heart)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    15 µg
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 14 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Aurora B antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human coronary artery endothelial cells HCAEC)
    Permeabilization
    Yes - 0.1% Trition X-100 in HBSS
    Specification
    Human coronary artery endothelial cells HCAEC
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 19 2020

    Western blot abreview for Anti-Aurora B antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human hepatocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    human hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 13 2017

    Western blot abreview for Anti-Aurora B antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hep G2)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Hep G2
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 13 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Aurora B antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Sheep Tissue sections (Fetal Heart)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer
    Permeabilization
    No
    Specification
    Fetal Heart
    Blocking step
    H2O2 & Histostain-Plus Blocking Solution as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 01 2017

    Western blot abreview for Anti-Aurora B antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Specification
    Cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 12 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Aurora B antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Cardiomyocytes)
    Permeabilization
    Yes - Triton x-100, 0.01%
    Specification
    Cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 12 2016

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Aurora B antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    Yes - Triton x-100, 0.01%
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 16 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Aurora B antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (heart)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: microwave
    Permeabilization
    Yes - 0.25% Triton
    Specification
    heart
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 20 2015

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