Overview

  • Product nameAnti-Ikaros antibody
    See all Ikaros primary antibodies
  • Description
    Rabbit polyclonal to Ikaros
  • Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chicken
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Ikaros.

    (Peptide available as ab35526.)

  • Positive control
    • This antibody gave a positive result in the following lysates: HeLa (Human Epithelial Carcinoma Cell Line) Whole Cell Lysata Jurkat (Human T cell Lymphoblast like Cell Line) Whole Cell Lysate Ramos (Human Burkitt's Lymphoma Cell Line) Whole Cell Lysate Spleen (Human) Tissue Lysate MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate This antibody gave a positive signal in the following Methanol fixed cell line: MCF-7.

Properties

Applications

Our Abpromise guarantee covers the use of ab26083 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 56 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionTranscription 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 specificityAbundantly expressed in thymus, spleen and peripheral blood Leukocytes and lymph nodes. Lower expression in bone marrow and small intestine.
  • Involvement in diseaseDefects 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 similaritiesBelongs to the Ikaros C2H2-type zinc-finger protein family.
    Contains 6 C2H2-type zinc fingers.
  • DomainThe 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 localizationCytoplasm; 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
  • FormThere 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

Anti-Ikaros antibody images

  • All lanes : Anti-Ikaros antibody (ab26083) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : Ramos (Human Burkitt's lymphoma cell line)Whole Cell Lysate
    Lane 4 : Spleen (Human) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 56 kDa
    Observed band size : 63 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 33 kDa (possible isoform).
  • IHC image of Ikaros staining in human tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab26083, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • Anti-Ikaros antibody (ab26083) at 1 µg/ml + MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 56 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab26083 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab26083 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Ikaros antibody (ab26083)

This product has been referenced in:
  • O'Brien S  et al. Ikaros imposes a barrier to CD8+ T cell differentiation by restricting autocrine IL-2 production. J Immunol 192:5118-29 (2014). Read more (PubMed: 24778448) »
  • Vitanza NA  et al. Ikaros deletions in BCR-ABL-negative childhood acute lymphoblastic leukemia are associated with a distinct gene expression signature but do not result in intrinsic chemoresistance. Pediatr Blood Cancer 61:1779-85 (2014). Read more (PubMed: 24976218) »

See all 5 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (C57BL/6 Mouse splenocytes)
Loading amount 40 µg
Specification C57BL/6 Mouse splenocytes
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Tomar Ghansah

Verified customer

Submitted Dec 14 2012

Thank you for your reply.

After checking the expression pattern of Ikaros, we would not expect this protein to be detectable in liver samples. As a positive control, I would recommend a spleen whole cell lysate prepared with RIPA buffer (such...

Read More

Thank you for contacting us. This antibody is validated using 5% BSA in 0.1% TBST as a blocking agent and antibody diluent, so it is possible that blocking with BSA for at least 1 hour at RT (up to overnight at 4C) may help to reduce the background sta...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (RS411 pre B leukemic cell line)
Loading amount 20 µg
Specification RS411 pre B leukemic cell line
Gel Running Conditions Reduced Denaturing (8%)
Blocking step 2% milk + 1% BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jan 23 2012

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (Nalm6 with Ty-1-IKAROS over expression)
Specification Nalm6 with Ty-1-IKAROS over expression
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Detection step Real-time PCR
Positive control Nalm6 transduced with Ty-1 IKAROS
Positive control for ChIP: Ty-1 antibody (MAb-054-050, Diagenode)
Negative control Parental Nalm6 cells
Username

Abcam user community

Verified customer

Submitted Jan 23 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"