Human Ikaros Antibody

Catalog # Availability Size / Price Qty
AF4984
AF4984-SP
Detection of Human Ikaros by Western Blot.
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Product Details
Citations (2)
FAQs
Supplemental Products
Reviews (1)

Human Ikaros Antibody Summary

Species Reactivity
Human
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.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human lkaros
Ser427-Ser519
Accession # Q13422
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Chromatin Immunoprecipitation (ChIP)
5 µg/5 x 106 cells
See below
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
Immersion fixed Jurkat human acute T cell leukemia cell line
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Ikaros antibody by Western Blot. View Larger

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.

Chromatin Immunoprecipitation (ChIP) Detection of Ikaros-regulated Genes antibody by Chromatin Immunoprecipitation. View Larger

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.

Intracellular Staining by Flow Cytometry Detection of Ikaros antibody in Jurkat Human Cell Line antibody by Flow Cytometry. View Larger

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.

Simple Western Detection of Human Ikaros antibody by Simple Western<SUP>TM</SUP>. View Larger

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.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

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
Entrez Gene IDs
10320 (Human); 22778 (Mouse); 305501 (Rat)
Alternate Names
DNA-binding protein Ikaros; hIk-1; IK1; IK1LyF-1; Ikaros (zinc finger protein); IKAROS family zinc finger 1 (Ikaros); Ikaros family zinc finger protein 1; Ikaros; IKAROSLymphoid transcription factor LyF-1; IKZF1; LYF1; LyF-1; LYF1PRO0758; PRO0758; zinc finger protein, subfamily 1A, 1 (Ikaros); ZNFN1A1; ZNFN1A1CLL-associated antigen KW-6

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Citations for Human Ikaros Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Immunomodulatory drugs activate NK cells via both Zap-70 and cereblon-dependent pathways
    Authors: T Hideshima, D Ogiya, J Liu, T Harada, K Kurata, J Bae, W Massefski, KC Anderson
    Leukemia, 2020-04-01;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. Epstein-Barr virus utilizes Ikaros in regulating its latent-lytic switch in B cells.
    Authors: Iempridee T, Reusch J, Riching A, Johannsen E, Dovat S, Kenney S, Mertz J
    J Virol, 2014-02-12;88(9):4811-27.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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Reviews for Human Ikaros Antibody

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Human Ikaros Antibody
By Magdalena Dragan on 01/20/2023
Application: Analysis of H929 whole cell lysates Sample Tested: Cell Lysates Species: Human