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  1. Link

    il-8-antibody-epr22994-255-ab235584.pdf

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Cardiovascular Angiogenesis Cytokines Interleukin
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IL-8 antibody [EPR22994-255] (ab235584)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (1)

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Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
  • Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
  • Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
  • Anti-IL-8 antibody [EPR22994-255] (ab235584)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22994-255] to IL-8
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
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Human CXCL8 knockout PC-3 cell line (ab273743)
Knockout
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Human CXCL8 knockout PC-3 cell lysate (ab275520)

View more associated products

Overview

  • Product name

    Anti-IL-8 antibody [EPR22994-255]
    See all IL-8 primary antibodies
  • Description

    Rabbit monoclonal [EPR22994-255] to IL-8
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: U-87 MG treated with 1µM Thapsigargin for 24 hours, whole cell lysate; Wild-type PC-3 LPS-treated (2 µg/ml, 6 h) with Brefeldin A (ab120299) (5 µg/ml, 5 h) cell lysate; His-tagged human recombiant IL-8 protein (aa21-99).
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22994-255
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Cytokines
    • Interleukin
    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-IL-8 antibody [EPR22994-255] - BSA and Azide free (ab275255)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • KO cell lines

    • Human CXCL8 knockout PC-3 cell line (ab273743)
  • KO cell lysates

    • Human CXCL8 knockout PC-3 cell lysate (ab275520)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab235584 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
1/1000. Predicted molecular weight: 11 kDa.
Notes
WB
1/1000. Predicted molecular weight: 11 kDa.
Application notes
Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.

Target

  • Function

    IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively.
  • Sequence similarities

    Belongs to the intercrine alpha (chemokine CxC) family.
  • Post-translational
    modifications

    Several N-terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukcocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8. IL-8(6-77) is the most promiment form.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P10145 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3576 Human
    • Omim: 146930 Human
    • SwissProt: P10145 Human
    • Unigene: 624 Human
    • Alternative names

      • (Ala-IL-8)77 antibody
      • (Ser-IL-8)72 antibody
      • 9E3 antibody
      • Beta thromboglobulin like protein antibody
      • C-X-C motif chemokine 8 antibody
      • CEF-4 antibody
      • chemokine, CXC motif, ligand 8 antibody
      • CXCL8 antibody
      • Emoctakin antibody
      • GCP-1 antibody
      • GCP/IL-8 protein I antibody
      • GCP/IL-8 protein II antibody
      • GCP/IL-8 protein III antibody
      • GCP/IL-8 protein IV antibody
      • GCP/IL-8 protein V antibody
      • GCP/IL-8 protein VI antibody
      • GCP1 antibody
      • Granulocyte chemotactic protein 1 antibody
      • IL-8 antibody
      • IL-8(1-77) antibody
      • IL-8(9-77) antibody
      • IL8 antibody
      • IL8/NAP1 form I antibody
      • IL8/NAP1 form II antibody
      • IL8/NAP1 form III antibody
      • IL8/NAP1 form IV antibody
      • IL8/NAP1 form V antibody
      • IL8/NAP1 form VI antibody
      • IL8_HUMAN antibody
      • Inteleukin 8 antibody
      • LECT antibody
      • LUCT antibody
      • Lymphocyte-derived neutrophil-activating factor antibody
      • LYNAP antibody
      • MDNCF antibody
      • MDNCF-b antibody
      • MDNCF-c antibody
      • MONAP antibody
      • Monocyte derived neutrophil activating peptide antibody
      • Monocyte derived neutrophil chemotactic factor antibody
      • Monocyte-derived neutrophil chemotactic factor antibody
      • Monocyte-derived neutrophil-activating peptide antibody
      • NAF antibody
      • NAP 1 antibody
      • NAP-1 antibody
      • NAP1 antibody
      • Neutrophil activating peptide 1 antibody
      • Neutrophil activating protein 1 antibody
      • Neutrophil-activating factor antibody
      • Neutrophil-activating protein 1 antibody
      • Protein 3 10C antibody
      • Protein 3-10C antibody
      • SCYB 8 antibody
      • SCYB8 antibody
      • Small inducible cytokine subfamily B member 8 antibody
      • T cell chemotactic factor antibody
      • T-cell chemotactic factor antibody
      • TSG 1 antibody
      • TSG1 antibody
      see all

    Images

    • Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
      Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
      All lanes : Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution

      Lane 1 : Wild-type PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate
      Lane 2 : Wild-type PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate
      Lane 3 : CXCL8 knockout PC-3 Brefeldin A (ab120299)-treated (5 μg/ml, 5 h) cell lysate
      Lane 4 : CXCL8 knockout PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (ab120299) (5 μg/ml, 5 h) cell lysate
      Lane 5 : A431 cell lysate
      Lane 6 : HCT116 cell lysate

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 11 kDa
      Observed band size: 10 kDa why is the actual band size different from the predicted?



      Lanes 1 - 6: Merged signal (red and green). Green - ab235584 observed at 10 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab235584 was shown to react with IL-8 in wild-type PC-3 cells in Western blot with loss of signal observed in CXCL8 knockout cell line ab273743 (knockout cell lysate ab275520). Wild-type PC-3 and CXCL8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab235584 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
      Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
      Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution + His-tagged human recombiant IL-8 protein (aa21-99) 10 ng

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 11 kDa
      Observed band size: 11 kDa



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      Exposure time: 3 minutes

       

    • Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
      Western blot - Anti-IL-8 antibody [EPR22994-255] (ab235584)
      All lanes : Anti-IL-8 antibody [EPR22994-255] (ab235584) at 1/1000 dilution

      Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
      Lane 2 : U-87 MG treated with 1µM Thapsigargin for 24 hours, whole cell lysate

      Lysates/proteins at 40 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 11 kDa
      Observed band size: 11 kDa



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      This blot was developed using a higher sensitivity ECL substrate.

      Exposure time: 3 minutes

       

    • Anti-IL-8 antibody [EPR22994-255] (ab235584)
      Anti-IL-8 antibody [EPR22994-255] (ab235584)

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab235584? Please let us know so that we can cite the reference in this datasheet.

    ab235584 has been referenced in 1 publication.

    • Zhao H & He Y The Inhibitory Effect of Lysophosphatidylcholine on Proangiogenesis of Human CD34+ Cells Derived Endothelial Progenitor Cells. Front Mol Biosci 8:682367 (2021). PubMed: 34179086

    Customer reviews and Q&As

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