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Human IL-6 knockout A549 cell lysate (ab275501)

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Western blot - Human IL-6 knockout A549 cell lysate (ab275501)
  • Western blot - Human IL6 knockout A549 cell lysate (ab275501)
  • Western blot - Human IL6 knockout A549 cell lysate (ab275501)
  • Sanger Sequencing - Human IL6 knockout A549 cell lysate

Overview

  • Product name

    Human IL-6 knockout A549 cell lysate
    See all IL-6 kits

  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

    Treatments:
    Human IL-6 knockout A549 cell lysate - Brefeldin A (5 µg/ml, 6h)
    Human wild-type A549 cell lysate - Brefeldin A (5 µg/ml, 6h)
    Human IL-6 knockout A549 cell lysate - IL-1ß (20 ng/ml, 24h) and Brefeldin A (5 µg/ml, during the last 6h of IL-1ß treatment)
    Human wild-type A549 cell lysate - IL-1ß (20 ng/ml, 24h) and Brefeldin A (5 µg/ml, during the last 6h of IL-1ß treatment)

  • Parental Cell Line

    A549

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 97 bp deletion in exon 3

  • Passage number

    <20

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)

  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

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.
  • Information by UniProt

  • Alternative names

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab275501 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
Use at an assay dependent concentration. Predicted molecular weight: 23 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 23 kDa.

Images

  • Western blot - Human IL-6 knockout A549 cell lysate (ab275501)
    Western blot - Human IL-6 knockout A549 cell lysate (ab275501)

    Lane 1: Wild-type A549 Vehicle Control IL-1b (0 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
    Lane 2: Wild-type A549 Treated IL-1b (20 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
    Lane 3: IL-6 knockout A549 Vehicle Control IL-1b (0 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
    Lane 4: IL-6 knockout A549 Treated IL-1b (20 ng/mL, 24 h), Brefeldin A (5 μg/mL, final 6 h) cell lysate 30 μg
    False colour image of Western blot: Anti-IL-6 antibody [EPR22565-204] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab233551 was shown to bind specifically to IL-6. A band was observed at 25 kDa in wild-type A549 cell lysates with no signal observed at this size in IL6 knockout cell line ab273751 (knockout cell lysate ab275501). To generate this image, wild-type and IL6 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Human IL6 knockout A549 cell lysate (ab275501)
    Western blot - Human IL6 knockout A549 cell lysate (ab275501)

    Lane 1: Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
    Lane 2: Wild-type A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
    Lane 3: IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
    Lane 4: IL-6 knockout A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
    Lanes 1 - 4: Merged signal (red and green). Green - ab233706 observed at 25 kDa, possible non-specific binding at 40 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.

  • Western blot - Human IL6 knockout A549 cell lysate (ab275501)
    Western blot - Human IL6 knockout A549 cell lysate (ab275501)

    Lane 1: Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
    Lane 2: Wild-type A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
    Lane 3: IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
    Lane 4: IL-6 knockout A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
    Lanes 1 - 4: Merged signal (red and green). Green - ab214429 observed at 25 kDa, possible non-specific binding at 35 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 A549 and IL-6 knockout 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.

  • Sanger Sequencing - Human IL6 knockout A549 cell lysate
    Sanger Sequencing - Human IL6 knockout A549 cell lysate

    Allele-1: 97 bp deletion in exon 3.

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

References (0)

Publishing research using ab275501? Please let us know so that we can cite the reference in this datasheet.

ab275501 has not yet been referenced specifically in any publications.

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