Ikaros/IKZF1 Antibody [Unconjugated]

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.

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. The VPAC promoter was detected by standard PCR.

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 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 the12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, ChIP, CyTOF-ready, Flow, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Ikaros/IKZF1 Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: IKZF1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • CyTOF-ready
  • Immunocytochemistry 5-15 ug/mL
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Ikaros/IKZF1 Antibody [Unconjugated]

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ikaros/IKZF1 Antibody (AF4984)(2)

Publications using AF4984

• T Hideshima, D Ogiya, J Liu, T Harada, K Kurata, J Bae, W Massefski, KC Anderson Immunomodulatory drugs activate NK cells via both Zap-70 and cereblon-dependent pathways Leukemia, 2020-04-01;0(0):. 2020-04-01 [PMID: 32238854] (Western Blot, Human)
• Iempridee T, Reusch J, Riching A, Johannsen E, Dovat S, Kenney S, Mertz J Epstein-Barr virus utilizes Ikaros in regulating its latent-lytic switch in B cells. J Virol, 2014-02-12;88(9):4811-27. 2014-02-12 [PMID: 24522918] (Western Blot, Human)

Bioinformatics

• Gene Symbol: IKZF1
• Uniprot:
  • Q13422()