Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, Chromatin Immunoprecipitation (ChIP), CyTOF-ready

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Ikaros Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human lkaros
Ser427-Ser519
Accession # Q13422

Specificity

Detects human lkaros in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross‑reactivity with recombinat human (rh) ZIC-1, rhZNF-24, and rhZNF-206 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Ikaros Antibody

Detection of Human Ikaros antibody by Western Blot.

Detection of Human Ikaros by Western Blot.

Western blot shows lysates of Nalm-6 human Pre-B acute lymphocytic leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Ikaros Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4984) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). Bands were detected for Ikaros (1-8 spice forms) at approximately 37 to 63 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Ikaros-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of Ikaros-regulated Genes by Chromatin Immunoprecipitation.

Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Ikaros/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human Ikaros Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4984) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. TheVPACpromoter was detected by standard PCR.

Detection of Ikaros antibody in Jurkat Human Cell Line antibody by Flow Cytometry.

Detection of Ikaros in Jurkat Human Cell Line by Flow Cytometry.

Jurkat human acute T cell leukemia cell line was stained with Goat Anti-Human Ikaros Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4984, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

Detection of Human Ikaros antibody by Simple WesternTM.

Detection of Human Ikaros by Simple WesternTM.

Simple Western lane view shows lysates of Nalm-6 human Pre-B acute lymphocytic leukemia cell line, loaded at 0.2 mg/mL. Specific bands were detected for Ikaros at approximately 63-77 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Ikaros Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4984) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human Ikaros Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: PMA and calcium ionomycin treated Jurkat human acute T cell leukemia cell line chromatin, VPAC promoter detected by standard PCR

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed Jurkat human acute T cell leukemia cell line

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line fixed with paraformaldehyde and permeabilized with methanol

Simple Western

10 µg/mL
Sample: Nalm‑6 human Pre-B acute lymphocytic leukemia cell line

Western Blot

1 µg/mL
Sample: Nalm‑6 human Pre‑B acute lymphocytic leukemia cell line

Reviewed Applications

Read 1 review rated 4 using AF4984 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Ikaros

Ikaros (also LyF1) is a 60 kDa member of the C2H2-type zinc-finger protein family. It is found in both T and B cells and serves as a context‑dependent activator or repressor of genes. Human Ikaros is 519 amino acids (aa) in length. It possesses two zinc‑finger domains, one at the N‑terminus that contains four zinc-finger motifs (aa 117‑224) and one at the C‑terminus that contains two zinc‑finger motifs (aa 462‑514). The C‑terminal motifs mediate homo- or heterodimerization, while the N‑terminal motifs bind DNA. Multiple splice forms of 37 kDa to 47 kDa exist (Ikaros 2‑8) with reduced numbers of N‑terminal zinc-finger motifs. At least three are needed for DNA binding, and a heterodimer with a two‑motif monomer is suggested to be transcriptionally inert. Over aa 427‑519 (which are not spliced), human Ikaros shows 88% aa identity to mouse Ikaros.

Long Name

DNA-binding protein Ikaros

Alternate Names

IK1, IKZF1, LyF-1, LYF1, PRO0758, ZNFN1A1

Entrez Gene IDs

10320 (Human); 22778 (Mouse); 305501 (Rat)

Gene Symbol

IKZF1

UniProt

Q13422

Additional Ikaros Products

Product Documents for Human Ikaros Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Ikaros Antibody

For research use only

Citations for Human Ikaros Antibody

Customer Reviews for Human Ikaros Antibody (1)

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  • Human Ikaros Antibody
    Name: Magdalena Dragan
    Application: Analysis of H929 whole cell lysates
    Sample Tested: Cell Lysates
    Species: Human
    Verified Customer | Posted 01/20/2023
    H929 whole cell lysates, 0.4mg/ml protein concentration, Anti Ikaros 1:50, Goat HRP 1:200
    Human Ikaros Antibody AF4984

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