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  1. Link

    il-6-antibody-epr21711-ab233706.pdf

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Immunology Innate Immunity Cytokines Interleukins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IL-6 antibody [EPR21711] (ab233706)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-IL-6 antibody [EPR21711] (ab233706)
  • Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR21711] (ab233706)
  • Flow Cytometry - Anti-IL-6 antibody [EPR21711] (ab233706)
  • Immunoprecipitation - Anti-IL-6 antibody [EPR21711] (ab233706)
  • Western blot - Anti-IL-6 antibody [EPR21711] (ab233706)
  • Anti-IL-6 antibody [EPR21711] (ab233706)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21711] to IL-6
  • Suitable for: WB, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

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Recombinant human IL-6 protein (Active) (ab259381)
Knockout
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Human IL-6 knockout A549 cell line (ab273751)

View more associated products

Overview

  • Product name

    Anti-IL-6 antibody [EPR21711]
    See all IL-6 primary antibodies
  • Description

    Rabbit monoclonal [EPR21711] to IL-6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human IL-6 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P05231

  • Positive control

    • WB: LPS/BFA-treated HUVEC whole cell lysate; Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate ICC/IF: LPS/BFA-treated HUVEC cells. Flow Cytometry: LPS/BFA-treated HUVEC cells. IP: LPS/BFA-treated HUVEC whole cell lysate.
  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21711
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Neuroscience
    • Neurology process
    • Metabolism
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Metabolic disorders

Associated products

  • Alternative Versions

    • Anti-IL-6 antibody [EPR21711] - BSA and Azide free (ab233707)
    • Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)
  • Conjugation kits

    • Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human IL-6 knockout A549 cell line (ab273751)
  • KO cell lysates

    • Human IL-6 knockout A549 cell lysate (ab275501)
  • Recombinant Protein

    • Recombinant human IL-6 protein (Active) (ab259381)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

Applications

Our Abpromise guarantee covers the use of ab233706 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. Detects a band of approximately 21 kDa (predicted molecular weight: 23 kDa).
ICC/IF 1/50.
Flow Cyt 1/500.
IP 1/30.

Target

  • Function

    Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoeitic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance.
  • Involvement in disease

    Genetic variations in IL6 are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis.
    Note=A IL6 promoter polymorphism is associated with a lifetime risk of development of Kaposi sarcoma in HIV-infected men.
  • Sequence similarities

    Belongs to the IL-6 superfamily.
  • Post-translational
    modifications

    N- and O-glycosylated.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P05231 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3569 Human
    • Omim: 147620 Human
    • SwissProt: P05231 Human
    • Unigene: 654458 Human
    • Alternative names

      • Interleukin BSF 2 antibody
      • B cell differentiation factor antibody
      • B cell stimulatory factor 2 antibody
      • B-cell stimulatory factor 2 antibody
      • BSF 2 antibody
      • BSF-2 antibody
      • BSF2 antibody
      • CDF antibody
      • CTL differentiation factor antibody
      • Cytotoxic T cell differentiation factor antibody
      • Hepatocyte stimulating factor antibody
      • Hepatocyte stimulatory factor antibody
      • HGF antibody
      • HSF antibody
      • Hybridoma growth factor antibody
      • Hybridoma growth factor Interferon beta-2 antibody
      • Hybridoma plasmacytoma growth factor antibody
      • IFN-beta-2 antibody
      • IFNB2 antibody
      • IL 6 antibody
      • IL-6 antibody
      • IL6 antibody
      • IL6_HUMAN antibody
      • Interferon beta 2 antibody
      • Interferon beta-2 antibody
      • Interleukin 6 antibody
      • Interleukin 6 (interferon beta 2) antibody
      • Interleukin BSF 2 antibody
      • Interleukin-6 antibody
      see all

    Images

    • Western blot - Anti-IL-6 antibody [EPR21711] (ab233706)
      Western blot - Anti-IL-6 antibody [EPR21711] (ab233706)
      All lanes : Anti-IL-6 antibody [EPR21711] (ab233706) at 1/1000 dilution

      Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
      Lane 2 : Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate
      Lane 3 : IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
      Lane 4 : IL-6 knockout A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 23 kDa
      Observed band size: 25 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 40 kDa (possible non-specific binding)



      Lanes 1 - 4: Merged signal (red and green). Green - ab233706 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

      ab233706 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab233706 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] 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 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR21711] (ab233706)
      Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR21711] (ab233706)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial cell line) cells labeling IL6 with ab233706 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HUVEC cells treated with lipopolysaccharide (0.5 μg/ml) for 24h and Brefeldin A (300 ng/ml) for 20h. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Flow Cytometry - Anti-IL-6 antibody [EPR21711] (ab233706)
      Flow Cytometry - Anti-IL-6 antibody [EPR21711] (ab233706)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HUVEC (human umbilical vein endothelial cell line) treated with lipopolysaccharide (0.5 μg/ml) for 24h and Brefeldin A (300 ng/ml) for 20h (red) / untreated control (green) cells  labeling IL6 with ab233706 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-IL-6 antibody [EPR21711] (ab233706)
      Immunoprecipitation - Anti-IL-6 antibody [EPR21711] (ab233706)

      IL6 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell line) treated with lipopolysaccharide (0.5 µg/ml) for 24h and Brefeldin A (300 ng/ml) for 20h, whole cell lysate with ab233706 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233706 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HUVEC treated with lipopolysaccharide (0.5 µg/ml) for 24h and Brefeldin A (300 ng/ml) for 20h, whole cell lysate 10 µg (Input).
      Lane 2: ab233706 IP in HUVEC treated with lipopolysaccharide (0.5 µg/ml) for 24h and Brefeldin A (300 ng/ml) for 20h, whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab233706 in HUVEC treated with lipopolysaccharide (0.5 µg/ml) for 24h and Brefeldin A (300 ng/ml) for 20h, whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 5 seconds.

    • Western blot - Anti-IL-6 antibody [EPR21711] (ab233706)
      Western blot - Anti-IL-6 antibody [EPR21711] (ab233706)
      All lanes : Anti-IL-6 antibody [EPR21711] (ab233706) at 1/1000 dilution

      Lane 1 : Untreated HUVEC (human umbilical vein endothelial cell line) whole cell lysate
      Lane 2 : HUVEC treated with 0.5 µg/ml lipopolysaccharides (LPS) for 24 hours, added 300 ng/ml BFA last 20 hours, whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 23 kDa
      Observed band size: 21 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/ Dilution buffer and concentration: 5% NFDM/TBST.

      The molecular mass observed is consistent with what has been described in the literature (PMID: 2523818, PMID: 2783321).

    • Anti-IL-6 antibody [EPR21711] (ab233706)
      Anti-IL-6 antibody [EPR21711] (ab233706)

    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

    • Datasheet

    Certificate of Compliance

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

  • References (2)

    Publishing research using ab233706? Please let us know so that we can cite the reference in this datasheet.

    ab233706 has been referenced in 2 publications.

    • Chen S  et al. Silencing CDC25A inhibits the proliferation of liver cancer cells by downregulating IL-6 in vitro and in vivo. Int J Mol Med 45:743-752 (2020). PubMed: 31922225
    • Hong S  et al. Sevoflurane induces apoptosis of isolated placental trophoblast cells and stimulates expressions of TNF-a and IL-6. Am J Transl Res 11:7084-7093 (2019). PubMed: 31814911

    Customer reviews and Q&As

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