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    products/primary-antibodies/il-6-antibody-epr20653-ab214429.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 [EPR20653] (ab214429)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (2)

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Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)
  • Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR20653] (ab214429)
  • Immunoprecipitation - Anti-IL-6 antibody [EPR20653] (ab214429)
  • Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)
  • Anti-IL-6 antibody [EPR20653] (ab214429)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20653] to IL-6
  • Suitable for: WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-IL-6 antibody [EPR20653]
    See all IL-6 primary antibodies
  • Description

    Rabbit monoclonal [EPR20653] to IL-6
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HUVEC treated with 0.5 µg/ml Lipopolysaccharides (LPS) for 24 hours and 300 ng/ml Brefeldin A (BFA) for 20 hours 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: HUVEC treated with Lipopolysaccharides (LPS) (0.5 µg/ml 24h) and Brefeldin A (BFA) (300 ng/ml 20h) cells. IP: HUVEC treated with 0.5 µg/ml Lipopolysaccharides (LPS) for 24 hours and 300 ng/ml Brefeldin A (BFA) for 20 hours 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.

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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20653
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Neuroscience
    • Neurology process
    • Metabolism
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Metabolic disorders
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-IL-6 antibody [EPR20653] - BSA and Azide free (ab245770)
  • 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

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab214429 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 21-28 kDa (predicted molecular weight: 23 kDa).
ICC/IF
1/500.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 21-28 kDa (predicted molecular weight: 23 kDa).
ICC/IF
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 [EPR20653] (ab214429)
      Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)
      All lanes : Anti-IL-6 antibody [EPR20653] (ab214429) 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: 35 kDa (possible non-specific binding)



      Lanes 1 - 4: Merged signal (red and green). Green - ab214429 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

      ab214429 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 ab214429 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 [EPR20653] (ab214429)
      Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR20653] (ab214429)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial cell line) cells, untreated or treated with Lipopolysaccharides (LPS) (0.5 µg/ml 24 hours) and Brefeldin A (BFA) (300 ng/ml 20 hours), labeling IL6 with ab214429 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing the cytoplasmic expression increased on HUVEC cells treated with Lipopolysaccharides (LPS) (0.5 µg/ml, 24 hours) and Brefeldin A (BFA) (300 ng/ml, 20 hours).

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      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.

    • Immunoprecipitation - Anti-IL-6 antibody [EPR20653] (ab214429)
      Immunoprecipitation - Anti-IL-6 antibody [EPR20653] (ab214429)

      IL6 was immunoprecipitated from 0.35 mg of HUVEC (human umbilical vein endothelial cell line) treated with Lipopolysaccharides (LPS) and Brefeldin A (BFA) whole cell lysate with ab214429 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214429 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

      Lane 1: HUVEC treated with 0.5 μg/ml LPS for 24 hours and 300 ng/ml BFA for 20 hours whole cell lysate 10 μg (Input). 

      Lane 2: ab214429 IP in HUVEC treated with 0.5 μg/ml LPS for 24 hours and 300 ng/ml BFA for 20 hours whole cell lysate (+). 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214429 in HUVEC treated with 0.5 μg/ml LPS for 24 hours and 300 ng/ml BFA for 20 hours whole cell lysate (-).

      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)
      Western blot - Anti-IL-6 antibody [EPR20653] (ab214429)
      All lanes : Anti-IL-6 antibody [EPR20653] (ab214429) 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 and 300 ng/ml Brefeldin A (BFA) for 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: 21-28 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The MW observed is consistent with the literature (PMID:2523818, PMID: 2783321).

    • Anti-IL-6 antibody [EPR20653] (ab214429)
      Anti-IL-6 antibody [EPR20653] (ab214429)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols

    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 (2)

    Publishing research using ab214429? Please let us know so that we can cite the reference in this datasheet.

    ab214429 has been referenced in 2 publications.

    • Dou B  et al. The roles of metastasis-related proteins in the development of giant cell tumor of bone, osteosarcoma and Ewing's sarcoma. Technol Health Care 29:91-101 (2021). PubMed: 33682749
    • Tobío A  et al. Oncogenic D816V-KIT signaling in mast cells causes persistent IL-6 production. Haematologica 105:124-135 (2020). PubMed: 30948489

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