Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Ikaros antibody [EPR13790] (ab191394)

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Western blot - Anti-Ikaros antibody [EPR13790] (ab191394)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (ab191394)
  • Western blot - Anti-Ikaros antibody [EPR13790] (ab191394)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (ab191394)
  • Flow Cytometry (Intracellular) - Anti-Ikaros antibody [EPR13790] (ab191394)
  • Anti-Ikaros antibody [EPR13790] (ab191394)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13790] to Ikaros
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Ikaros antibody [EPR13790]
    See all Ikaros primary antibodies

  • Description

    Rabbit monoclonal [EPR13790] to Ikaros

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Jurkat, Daudi, Raji, Ramos and MOLT4 cell lysates; Human thymus and mouse spleen tissues; MOLT4 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab191394 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
Flow Cyt (Intra)
1/130.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/10000 - 1/50000. Detects a band of approximately 50-70 kDa (predicted molecular weight: 58 kDa).
IHC-P
1/1400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
1/130.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/10000 - 1/50000. Detects a band of approximately 50-70 kDa (predicted molecular weight: 58 kDa).
IHC-P
1/1400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Transcription regulator of hematopoietic cell differentiation (PubMed:17934067). Binds gamma-satellite DNA (PubMed:17135265, PubMed:19141594). Plays a role in the development of lymphocytes, B- and T-cells. Binds and activates the enhancer (delta-A element) of the CD3-delta gene. Repressor of the TDT (fikzfterminal deoxynucleotidyltransferase) gene during thymocyte differentiation. Regulates transcription through association with both HDAC-dependent and HDAC-independent complexes. Targets the 2 chromatin-remodeling complexes, NuRD and BAF (SWI/SNF), in a single complex (PYR complex), to the beta-globin locus in adult erythrocytes. Increases normal apoptosis in adult erythroid cells. Confers early temporal competence to retinal progenitor cells (RPCs) (By similarity). Function is isoform-specific and is modulated by dominant-negative inactive isoforms (PubMed:17135265, PubMed:17934067).

  • Tissue specificity

    Abundantly expressed in thymus, spleen and peripheral blood Leukocytes and lymph nodes. Lower expression in bone marrow and small intestine.

  • Involvement in disease

    Defects in IKZF1 are frequent occurrences (28.6%) in acute lymphoblasic leukemia (ALL). Such alterations or deletions lead to poor prognosis for ALL.
    Chromosomal aberrations involving IKZF1 are a cause of B-cell non-Hodgkin lymphomas (B-cell NHL). Translocation t(3;7)(q27;p12), with BCL6.

  • Sequence similarities

    Belongs to the Ikaros C2H2-type zinc-finger protein family.
    Contains 6 C2H2-type zinc fingers.

  • Domain

    The N-terminal zinc-fingers 2 and 3 are required for DNA binding as well as for targeting IKFZ1 to pericentromeric heterochromatin.
    The C-terminal zinc-finger domain is required for dimerization.

  • Post-translational
    modifications

    Phosphorylation controls cell-cycle progression from late G(1) stage to S stage. Hyperphosphorylated during G2/M phase. Dephosphorylated state during late G(1) phase. Phosphorylation on Thr-140 is required for DNA and pericentromeric location during mitosis. CK2 is the main kinase, in vitro. GSK3 and CDK may also contribute to phosphorylation of the C-terminal serine and threonine residues. Phosphorylation on these C-terminal residues reduces the DNA-binding ability. Phosphorylation/dephosphorylation events on Ser-13 and Ser-295 regulate TDT expression during thymocyte differentiation. Dephosphorylation by protein phosphatase 1 regulates stability and pericentromeric heterochromatin location. Phosphorylated in both lymphoid and non-lymphoid tissues (By similarity). Phosphorylation at Ser-361 and Ser-364 downstream of SYK induces nuclear translocation.
    Sumoylated. Simulataneous sumoylation on the 2 sites results in a loss of both HDAC-dependent and HDAC-independent repression. Has no effect on pericentromeric heterochromatin location. Desumoylated by SENP1.
    Polyubiquitinated.

  • Cellular localization

    Cytoplasm; Nucleus. In resting lymphocytes, distributed diffusely throughout the nucleus. Localizes to pericentromeric heterochromatin in proliferating cells. This localization requires DNA binding which is regulated by phosphorylation / dephosphorylation events and Nucleus. In resting lymphocytes, distributed diffusely throughout the nucleus. Localizes to pericentromeric heterochromatin in proliferating cells. This localization requires DNA binding which is regulated by phosphorylation / dephosphorylation events (By similarity).
  • Information by UniProt

  • Database links

    see all

  • Form

    There are 7 isoforms produced by alternative splicing.

  • Alternative names

    • CLL associated antigen KW 6 antibody
    • DNA-binding protein Ikaros antibody
    • hIk 1 antibody
    • Hs.54452 antibody
    • IK1 antibody
    • Ikaros (zinc finger protein) antibody
    • IKAROS antibody
    • IKAROS family zinc finger 1 (Ikaros) antibody
    • Ikaros family zinc finger protein 1 antibody
    • Ikzf1 antibody
    • IKZF1_HUMAN antibody
    • LYF1 antibody
    • Lymphoid transcription factor LyF-1 antibody
    • PRO0758 antibody
    • Zinc finger protein subfamily 1A 1 (Ikaros) antibody
    • Zinc finger protein subfamily 1A 1 antibody
    • Zinc finger protein, subfamily 1A, member 1 antibody
    • ZNFN1A1 antibody
    see all

Images

  • Western blot - Anti-Ikaros antibody [EPR13790] (ab191394)
    Western blot - Anti-Ikaros antibody [EPR13790] (ab191394)
    All lanes : Anti-Ikaros antibody [EPR13790] (ab191394) at 1/10000 dilution

    Lane 1 : Wild-type Jurkat cell lysate
    Lane 2 : IKZF1 knockout Jurkat cell lysate
    Lane 3 : Daudi cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 50-70 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab191394 observed at 50-70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab191394 was shown to react with Ikaros in wild-type Jurkat cells in western blot with loss of signal observed in IKZF1 knockout sample. Wild-type and IKZF1 knockout Jurkat cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab191394 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (ab191394)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (ab191394)

    Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling Ikaros with ab191394 at 1/1400 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.
    Inset: Negative control: using PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Ikaros antibody [EPR13790] (ab191394)
    Western blot - Anti-Ikaros antibody [EPR13790] (ab191394)
    All lanes : Anti-Ikaros antibody [EPR13790] (ab191394) at 1/10000 dilution

    Lane 1 : Raji cell lysate
    Lane 2 : Ramos cell lysate
    Lane 3 : MOLT4 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 58 kDa



    Based on the sequence analysis, ab191394 recognizes seven isoforms with the predicted MWs of 58KDa, 48KDa, 48KDa, 43KDa, 41KDa, 32KDa and 53KDa, respectively.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (ab191394)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (ab191394)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Ikaros with ab191394 at 1/1400 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.
    Inset: Negative control: using PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-Ikaros antibody [EPR13790] (ab191394)
    Flow Cytometry (Intracellular) - Anti-Ikaros antibody [EPR13790] (ab191394)

    Intracellular flow cytometric analysis of MOLT4 cells (paraformaldehyde-fixed, 2%)labeling Ikaros with ab191394 at 1/130 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution. 

     

     

  • Anti-Ikaros antibody [EPR13790] (ab191394)
    Anti-Ikaros antibody [EPR13790] (ab191394)

References (2)

Publishing research using ab191394? Please let us know so that we can cite the reference in this datasheet.

ab191394 has been referenced in 2 publications.

  • Liu J  et al. Light-induced control of protein destruction by opto-PROTAC. Sci Adv 6:eaay5154 (2020). PubMed: 32128407
  • Boutboul D  et al. Dominant-negative IKZF1 mutations cause a T, B, and myeloid cell combined immunodeficiency. J Clin Invest 128:3071-3087 (2018). PubMed: 29889099

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