Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)

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 - Low endotoxin, Azide free
  • Suitable for: WB, IP, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free
    See all IL-6 primary antibodies

  • Description

    Rabbit monoclonal [EPR21711] to IL-6 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: LPS/BFA-treated HUVEC whole cell lysate; Wild-type A549 IL-1b (ab259387)-treated (20 ng/ml, 24h) and Brefeldin A (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

    ab246703 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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

Images

  • Western blot - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)
    Western blot - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)
    All lanes : Anti-IL-6 antibody [EPR21711] (ab233706) at 1/1000 dilution

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

    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)



    This data was developed using the same antibody clone in a different buffer formulation (ab233706).

    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] - Low endotoxin, Azide free (ab246703)
    Immunocytochemistry/ Immunofluorescence - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab233706).

  • Flow Cytometry - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)
    Flow Cytometry - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab233706).

  • Immunoprecipitation - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)
    Immunoprecipitation - Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab233706).

  • Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)
    Anti-IL-6 antibody [EPR21711] - Low endotoxin, Azide free (ab246703)

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