Overview

  • Product name

    Anti-IKZF3 antibody [EPR9342(B)]
    See all IKZF3 primary antibodies
  • Description

    Rabbit monoclonal [EPR9342(B)] to IKZF3
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human IKZF3 aa 1-100 (N terminal). The exact sequence is proprietary.
    (Peptide available as ab184024)

  • Positive control

  • 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 ab139408 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 1/20000. Predicted molecular weight: 58 kDa.Can be blocked with IKZF3 peptide (ab184024).

For unpurified use at 1/1000 - 1/10000.

IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

ICC/IF 1/100.

For unpurified use at 1/250 - 1/500.

Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Transcription factor that plays an important role in the regulation of lymphocyte differentiation. Plays an essential role in regulation of B-cell differentiation, proliferation and maturation to an effector state. Involved in regulating BCL2 expression and controlling apoptosis in T-cells in an IL2-dependent manner.
  • Tissue specificity

    Expressed most strongly in peripheral blood leukocytes, the spleen, and the thymus.
  • Sequence similarities

    Belongs to the Ikaros C2H2-type zinc-finger protein family.
    Contains 6 C2H2-type zinc fingers.
  • Post-translational
    modifications

    Phosphorylation on tyrosine residues induced by IL2 is required for dissociation from HRAS and nuclear translocation of IKZF3 in T-cells. Phosphorylation on tyrosine residues induced by IL4 is required for dissociation from Bcl-X(L) in T-cells.
  • Cellular localization

    Nucleus. Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • AIO antibody
    • Aiolos antibody
    • IKAROS family zinc finger 3 (Aiolos) antibody
    • IKAROS family zinc finger 3 antibody
    • Ikaros family zinc finger protein 3 antibody
    • IKZF 3 antibody
    • IKZF3 antibody
    • IKZF3_HUMAN antibody
    • zinc finger DNA binding protein Aiolos antibody
    • Zinc finger protein Aiolos antibody
    • Zinc finger protein subfamily 1A 3 (Aiolos) antibody
    • Zinc finger protein subfamily 1A 3 antibody
    • Zinc finger protein subfamily 1A, member 3 antibody
    • ZNFN1A3 antibody
    see all

Images

  • All lanes : Anti-IKZF3 antibody [EPR9342(B)] (ab139408) at 1/20000 dilution (purified)

    Lane 1 : Raji (Human Burkitt's lymphoma cell line) cell lysate
    Lane 2 : Ramos (Human Burkitt's lymphoma cell line) cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 58 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?



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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue labeling IKZF3 with purified ab139408 at 1/100.

    Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

    Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling IKZF3 (green) with purified ab139408 at 1/100.

    Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: Secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • Flow cytometry analysis of Ramos (Human Burkitt's lymphoma cell line cells labeling IKZF3 with purified ab139408 at 1/90 (red).

    Cells were fixed with 2% paraformaldehyde. An FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.

  • All lanes : Anti-IKZF3 antibody [EPR9342(B)] (ab139408) at 1/1000 dilution (unpurified)

    Lane 1 : Human thymus cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 3 : Raji (Human Burkitt's lymphoma cell line) cell lysate
    Lane 4 : Ramos (Human Burkitt's lymphoma cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 58 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labeling IKZF3 with purified ab139408 at 1/100.

    Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

    Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue labeling IKZF3 with purified ab139408 at 1/100.

    Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

    Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labeling IKZF3 with unpurified ab139408 at 1/250 dilution.

References

This product has been referenced in:

See all 6 Publications for this product

Customer reviews and Q&As

Answer

Thank you for your inquiry.

This is the intracellular flow cytometry protocol that was used with ab139408:

Flow Cytometry Protocol for Intracellular Staining

1. Solutions and Reagents
1.1. 1X PBS

1.2. Blocking buffer: 0.5% BSA in 1X PBS

1.3. 2% paraformaldehyde (1% solution - optional for storing samples)

1.4. 1X FACS permeabilizing solution (BD Biosciences cat. #340973)

1.5. Fluorescently-conjugated secondary antibody (various forms)

2. Protocol

2.1. Collect 1x10^6 cells/sample.

2.2. Wash cells once with blocking buffer.

2.3. Fix cells with 2% paraformaldehyde and incubate at room temperature for 10 min.

2.4. Wash cells once with blocking buffer.

2.5. Add 0.5 ml 1X FACS permeabilizing solution and incubate at room temperature for 10 min.

2.6. Wash cells once with blocking buffer.

2.7. Incubate cells in blocking buffer for 30 min at room temperature.

2.8. Add primary antibody at the appropriate dilution and incubate for 30 min at room temperature.

2.9. Wash twice with blocking buffer and incubate with fluorescently-conjugated anti-rabbit secondary antibody for 30 min at room temperature.

2.10. Wash cells twice with blocking buffer.

2.11. Re-suspend cells in 1X PBS and analyze on flow cytometry. Samples can be kept in 1% paraformaldehyde at 4 °C overnight.

I hope this information helps. Please contact us with any other questions.

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