Recombinant
RabMAb

Recombinant Anti-DLL3 antibody [EPR22592-18] (ab229902)

Overview

  • Product name

    Anti-DLL3 antibody [EPR22592-18]
    See all DLL3 primary antibodies
  • Description

    Rabbit monoclonal [EPR22592-18] to DLL3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human DLL3 aa 150-250. The exact sequence is proprietary.
    Database link: Q9NYJ7

  • Positive control

    • WB: HEK-293T transfected with DLL3 (WT) expression vector containing a myc-His-tag®, whole cell lysate, TT cell lysate. IHC-P: Human small cell lung cancer, cerebrum and glioma tissue; Mouse embryonic brain of day 14.5 tissue. ICC/IF: HEK-293T cells. Flow: HEK-293T cells. IP: HEK-293T transfected with DLL3 expression construct containing a myc-His-tag® whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab229902 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 75 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/600.
IP 1/30.

Target

  • Function

    Inhibits primary neurogenesis. May be required to divert neurons along a specific differentiation pathway. Plays a role in the formation of somite boundaries during segmentation of the paraxial mesoderm.
  • Involvement in disease

    Spondylocostal dysostosis 1
  • Sequence similarities

    Contains 1 DSL domain.
    Contains 6 EGF-like domains.
  • Domain

    The 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
  • Database links

  • Alternative names

    • Delta Drosophila like 3 antibody
    • Delta like 3 Drosophila antibody
    • Delta like 3 homolog Drosophila antibody
    • Delta like 3 protein antibody
    • Delta like protein 3 precursor antibody
    • Delta-like protein 3 antibody
    • Delta3 antibody
    • Dll3 antibody
    • DLL3_HUMAN antibody
    • Drosophila Delta homolog 3 antibody
    • SCDO1 antibody
    • SCOD1 antibody
    • Spondylocostal dysostosis autosomal recessive antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human small cell lung cancer tissue labeling DLL3 with ab229902 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human small cell lung cancer (PMID: 28487384, 30397180) is observed.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab229902 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Anti-DLL3 antibody [EPR22592-18] (ab229902) at 1/1000 dilution + TT (human thyroid carcinoma epithelial cell) whole cell lysate at 10 µg with 5% NFDM/TBST

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 65 kDa
    Observed band size: 65 kDa

  • Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling DLL3 with ab229902 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human glioma (PMID: 30397180) is observed.

    The section was incubated with ab229902 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemical analysis of paraffin-embedded mouse embryonic brain of day 14.5 tissue labeling DLL3 with ab229902 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse embryonic brain of day 14.5 (PMID: 19389376) is observed.

    The section was incubated with ab229902 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling DLL3 with ab229902 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on human cerebrum (PMID: 30397180) is observed.

    The section was incubated with ab229902 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling DLL3 with ab229902 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on mouse cerebrum (PMID: 30397180) is observed.

    The section was incubated with ab229902 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with DLL3 expression construct containing a myc-His-tag®  labelling DLL3 with ab229902 at 1/600 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730, Left) isotype control. 

    Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Cells were stained with rabbit IgG (Left) or ab229902 (Right). Then stained with anti-myc-tag conjugated to Alexa Fluor® 647.

  • Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells labelling DLL3 with ab229902 at 1/100 dilution, followed by ab150080 AlexaFluor®594 Goat anti-Rabbit secondary antibody at 1/500 dilution (Green). An anti-myc-tag mAb (Alexa Fluor® 488 Conjugate) was used at 1/100 dilution. Confocal image showing positive staining in HEK-293T cells transfected with DLL3 expression construct containing a myc-His-tag® is observed.  The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150080 AlexaFluor®594 Goat anti-Rabbit secondary at 1/500 dilution.

  • DLL3 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) transfected with DLL3 expression construct containing a myc-His-tag®  whole cell lysate with ab229902 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab229902 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

    Lane 1: HEK-293T (human embryonic kidney epithelial cell) transfected with DLL3 expression construct containing a myc-His-tag®  whole cell lysate 10μg

    Lane 2: ab229902 IP in HEK-293T transfected with DLL3 expression construct containing a myc-His-tag®  whole cell lysate (Input).

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229902 in HEK-293T transfected with DLL3 expression construct containing a myc-His-tag®  whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 15 seconds. 

  • All lanes : Anti-DLL3 antibody [EPR22592-18] (ab229902) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DLL3 (WT) expression vector containing a myc-His-tag®, whole cell lysate
    Lane 2 : HEK-293T transfected with empty vector containing myc-His-tag®, 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

    Predicted band size: 65 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

References

ab229902 has not yet been referenced specifically in any publications.

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