Bioactive grade

Recombinant human DLL4 protein (Fc Chimera Active) (ab108557)

Description

  • Product name

    Recombinant human DLL4 protein (Fc Chimera Active)
    See all DLL4 proteins and peptides
  • Biological activity

    Inhibits adipogenesis of 3T3L-1 cells and mesenchymal stem cells (MSCs). Induces the Notch target gene HES-1 when coated on a plate at 1µg/ml.

  • Purity

    >= 95 % SDS-PAGE.

  • Endotoxin level

    < 0.010 Eu/µg
  • Expression system

    HEK 293 cells
  • Accession

  • Protein length

    Protein fragment
  • Animal free

    No
  • Nature

    Recombinant
    • Species

      Human
    • Predicted molecular weight

      80 kDa including tags
    • Amino acids

      1 to 529
    • Tags

      Fc tag C-Terminus
    • Additional sequence information

      Fused to the Fc region of Human IgG1 at the C-terminus.

Associated products

Specifications

Our Abpromise guarantee covers the use of ab108557 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    Functional Studies

    SDS-PAGE

  • Form

    Lyophilised
  • Additional notes

     ab108557 interacts with Human Notch1 (as confirmed by Flow Cytometry).

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

    Constituents: PBS, 0.5% Trehalose

    ab108557 is 0.2 µm filtered.

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • Reconstitution
    Reconstitute with 100µl sterile water. PBS containing at least 0.1% BSA should be used for further dilutions. Working aliquots are stable for up to 3 months when stored at -20°C.

General Info

  • Alternative names

    • AOS6
    • Delta 4
    • delta 4 precursor
    • Delta ligand 4
    • delta ligand 4 precursor
    • Delta like 4
    • Delta like 4 homolog
    • Delta like 4 protein
    • Delta like canonical Notch ligand 4
    • Delta like protein 4
    • Delta-like 4 (Drosophila)
    • Delta-like protein 4
    • Delta4
    • DLL 4
    • Dll4
    • DLL4_HUMAN
    • Drosophila Delta homolog 4
    • hdelta2
    • Homeobox protein DLL-4
    • MGC126344
    • Notch ligand delta 2
    • Notch ligand DLL4
    • Notch ligand DLL4 precursor
    • XDLL-4
    see all
  • Function

    Plays a role in the Notch signaling pathway. Activates Notch-1 and Notch-4.
  • Tissue specificity

    Expressed in vascular endothelium.
  • Sequence similarities

    Contains 1 DSL domain.
    Contains 8 EGF-like domains.
  • Domain

    The Delta-Serrate-Lag2 (DSL) domain is required for binding to the Notch receptor.
  • Post-translational
    modifications

    Ubiquitinated by MIB (MIB1 or MIB2), leading to its endocytosis and subsequent degradation.
  • Cellular localization

    Membrane.
  • Information by UniProt

Images

  • Interaction of Human Notch1 with Human DLL4.
    HEK293 cells transfected with a Human Notch1 or a Human GITR ligand expressing vector were incubated with 25 μg/ml of Human GITR-Fc or ab108557. Cells were stained with anti-Human IgG (Fc specific) FITC conjugate for DLL4-Fc binding.
  • Induction of Hes-1 with the treatment of recombinant Human DLL4-Fc (ab108557).


    A Mouse preadpipocyte cell line, 3T3L-1, was stimulated with 5 μg/ml of Human DLL4-Fc as in indicated time points and each cell lysate was prepared and subjected to western blot by using anti-Mouse Hes1 or GAPDH.

    M: Marker.
    Lane 1: hDLL4-Fc, 0 min.
    Lane 2: hDLL4-Fc, 10 min.
    Lane 3: hDLL4-Fc, 30 min.
    Lane 4: hDLL4-Fc, 1 hr.
    Lane 5: hDLL4-Fc, 2 hr.
    Lane 6: hDLL4-Fc, 4 hr.
    Lane 7: hDLL4-Fc, 8 hr.
    Lane 8: hDLL4-Fc, 24 hr.

  • Adipogenesis inhibition of 3T3L-1 cells.
    3T3L-1 cells (mouse pre-adipocyte cells) were maintained in DMEM, supplemented with 10% fetal bovine serum and penicillin-streptomycin. For differentiation of 3T3L-1 cells, 3T3L-1 cells were cultured in adipogenic medium which was growth medium supplemented with 1 μM Dexamethasone, 0.5 mM IBMX, 10 μg/ml lnsulin (day 0). Medium was changed every 2 days. Staining with Oil Red O was typically performed on day 7. Cells were washed twice with PBS, fixed with 3.7% formalin, and stained with 0.5% filtered Oil Red O in propylene glycol. For negative controls, mouse TNF-α (20 ng/ml) was added. Recombinant Human DLL4-Fc (ab108557) (5 μg/ml) dissolved in DPBS was added to the differentiation medium. These plates were then used to differentiate 3T3L-1 cells.
  • Adipogenesis inhibition of MSCs.
    MSCs (Mesenchymal stem cells) were maintained in DMEM, supplemented with 10% fetal bovine serum, penicilinstreptomycin and glutamine. For differentiation of MSCs, MSCs were cultured in adipogenic medium which was growth medium supplemented with 1 μM Dexamethasone, 0.5mM IBMX, 10 μg/m lnsulin, 100 μM Indomethacin (day 1). Medium was changed every 3 days. Staining with Oil Red O was typically performed on day 30. For negative controls, TNF-α (20 ng/ml) was added. To immobilize Notch ligands on the plastic surface of the culture plates, plates were incubated with a solution of ab108557 (5 μg/ml) or mCD137-Fc (5 μg/ml) in PBS for 2 hours at 37°C. Plates were then used to differentiate MSCs.
  • Adipogenesis inhibition of 3T3L-1 cells.
  • 50 μg of cell lysates derived from hDLL4-Fc (ab108557) or non-treated 3T3L-1 cells, which had been either differentiated or undifferentiated, and were subjected to Western blot by using a Mouse adiponectin antibody.

References

This product has been referenced in:

  • Kamalakar A  et al. A non-canonical JAGGED1 signal to JAK2 mediates osteoblast commitment in cranial neural crest cells. Cell Signal 54:130-138 (2019). Read more (PubMed: 30529759) »
  • Tiemeijer LA  et al. Spatial patterning of the Notch ligand Dll4 controls endothelial sprouting in vitro. Sci Rep 8:6392 (2018). Read more (PubMed: 29686270) »
See all 5 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for your reply.

I am confident that the reference ab90285 is a suitable negative control for ab108557.
Furthermore, I have just been informed about the price of the new Fc fragments which is $424.00 for 100µg, therefore I would recommend to keep using the much cheaper Fc fragments reference ab90285, $269.00 for 1mg.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

I would like to inform you that the originator of the DLL4 fused to the Fc portion of Human IgG1 (ab108557) is able to supply the Fc portion only.

Could you please let me know if you would still be interested in such product? If so we will add it to our catalog.

Read More

Answer

Thank you very much for contacting us.

I just wanted to let you know that I am still working on your inquiry.

I have contacted the originator of the DLL4 protein ab108557 who informed me thatthe Human IgG Fc fragment we have in our catalogue (ab90285, https://www.abcam.com/ab90285)could be usedas a negative control. However, we do not know what the endotoxin content of this product is and I am investigating this.

Iam very sorry for the delay. I will forward to you the information as soon as I receive it. Thank you for your patience.

Read More

Answer

Thank you very much for contacting us with your question.

I've looked over both of these products, and they are both active DLL4 proteins. I'm not sure why ab108557 does not have "active" in its name, but I will recommend that this is updated.

Ab108557 has been shown to inhibit adipogenesis of 3T3L-1 cells and MSC's and also to induce Hes-1 in 3T3L-1 cells.Ab84081 has been used to inhibit myogenesis in C2C12 cells.

The major difference between these proteins is that ab84081 corresponds to amino acids 27-524 of human DLL4, while ab108557 is amino acids 1-529 and it is fused to a human IgG1 Fc fragment at the C terminus.

I hope that this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

Read More

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