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    products/cell-lysates/human-tgfb1-tgf-beta-1-knockout-a549-cell-lysate-ab269671.pdf

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Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)

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Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)
  • Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)

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Overview

  • Product name

    Human TGFB1 (TGF beta 1) knockout A549 cell lysate
    See all TGF beta 1 kits
  • Product overview

     Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    A549
  • Organism

    Human
  • Mutation description

    Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 99.45%
  • Passage number

    <20
  • Knockout validation

    Next Generation Sequencing (NGS), 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab280546 - Human TGFB1 knockout A549cell lysate 1 x 100µg
    ab259782 - Human wild-type A549 cell lysate 1 x 100µg
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Growth Factors
    • TGF
    • Signal Transduction
    • Growth Factors/Hormones
    • TGF
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Secreted
    • Cancer
    • Growth factors
    • TGF
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Cofactors, Vitamins / minerals
    • Co-factors
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
  • Cell type

    epithelial
  • Disease

    Carcinoma
  • Gender

    Male

Target

  • Function

    Multifunctional protein that controls proliferation, differentiation and other functions in many cell types. Many cells synthesize TGFB1 and have specific receptors for it. It positively and negatively regulates many other growth factors. It plays an important role in bone remodeling as it is a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts.
  • Tissue specificity

    Highly expressed in bone. Abundantly expressed in articular cartilage and chondrocytes and is increased in osteoarthritis (OA). Co-localizes with ASPN in chondrocytes within OA lesions of articular cartilage.
  • Involvement in disease

    Defects in TGFB1 are the cause of Camurati-Engelmann disease (CE) [MIM:131300]; also known as progressive diaphyseal dysplasia 1 (DPD1). CE is an autosomal dominant disorder characterized by hyperostosis and sclerosis of the diaphyses of long bones. The disease typically presents in early childhood with pain, muscular weakness and waddling gait, and in some cases other features such as exophthalmos, facial paralysis, hearing difficulties and loss of vision.
  • Sequence similarities

    Belongs to the TGF-beta family.
  • Post-translational
    modifications

    Glycosylated.
    The precursor is cleaved into mature TGF-beta-1 and LAP, which remains non-covalently linked to mature TGF-beta-1 rendering it inactive.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix.
  • Target information above from: UniProt accession P01137 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Cartilage-inducing factor
    • CED
    • Differentiation inhibiting factor
    • DPD1
    • LAP
    • Latency-associated peptide
    • Prepro transforming growth factor beta 1
    • TGF beta
    • TGF beta 1
    • TGF beta 1 protein
    • TGF-beta 1 protein
    • TGF-beta-1
    • TGF-beta-5
    • TGF-beta1
    • TGFB
    • Tgfb-1
    • tgfb1
    • TGFB1_HUMAN
    • TGFbeta
    • TGFbeta1
    • Transforming Growth Factor b1
    • Transforming Growth Factor beta 1
    • Transforming growth factor beta 1a
    • transforming growth factor beta-1
    • transforming growth factor, beta 1
    • Transforming Growth Factor-ß1
    see all

Associated products

  • KO cell lines

    • Human TGFB1 (TGF beta 1) knockout A549 cell line (ab269509)
  • Related Products

    • Anti-TGF beta 1 antibody [EPR18163] (ab179695)
    • Anti-TGF beta 1 antibody [EPR21143] (ab215715)
    • Anti-TGF beta 1 antibody [EPR21143] - BSA and Azide free (ab229856)
    • Anti-TGF beta 1 antibody [EPR18163] - BSA and Azide free (ab233730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab269671 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.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)
    Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)

    Lane 1: Wild-type A549 cell lysate 20 μg
    Lane 2: TGFB1 knockout A549 cell lysate 20 μg
    Lane 3: K562 cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg

    Lanes 1 - 4: Merged signal (red and green). Green - ab215715 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab215715 was shown to react with TGF beta 1 in wild-type A549 cells in Western blot with loss of signal observed in TGFB1 knockout cell line ab269509 (TGFB1 knockout cell lysate ab269671). Wild-type A549 and TGFB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab215715 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 h at room temperature before imaging.

  • Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)
    Western blot - Human TGFB1 (TGF beta 1) knockout A549 cell lysate (ab269671)

    Lane 1: Wild-type A549 cell lysate 20 μg
    Lane 2: TGFB1 knockout A549 cell lysate 20 μg
    Lane 3: K562 cell lysate 20 μg
    Lane 4: SH-SY5Y cell lysate 20 μg

    Lanes 1 - 4: Merged signal (red and green). Green - ab179695 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab179695 was shown to react with TGF beta  in wild-type A549 cells in Western blot with loss of signal observed in TGFB1 knockout cell line ab269509 (TGFB1 knockout cell lysate ab269671). Wild-type A549 and TGFB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab179695 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 h at room temperature before imaging.

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

References (0)

Publishing research using ab269671? Please let us know so that we can cite the reference in this datasheet.

ab269671 has not yet been referenced specifically in any publications.

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