| 货号 | AF4984-SP |
| 别名 | 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; IKAROSLymphoid transcription factor LyF-1; IKZF1; LYF1; LyF-1; LYF1PRO0758; PRO0758; zinc finger protein, subfamily 1A, 1 (Ikaros); ZNFN1A1; ZNFN1A1CLL-associated antigen KW-6 | 全称 | DNA-binding protein Ikaros |
| 反应种属 | Human |
| 应用 | Western Blot,Simple Western,Chromatin Immunoprecipitation (ChIP),CyTOF-ready,Immunocytochemistry,Intracellular Staining by Flow Cytometry |
| 目标/特异性 | 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. |
| 使用方法 | Western Blot: 1 µg/mL Simple Western: 10 µg/mL Chromatin Immunoprecipitation (ChIP): 5 µg/5 x 106cells Immunocytochemistry: 5-15 µg/mL Intracellular Staining by Flow Cytometry: 2.5 µg/106cells |
| 来源 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 产品组分 |
| 供应商 | R&D Systems |
| Entrez Gene IDs | 10320 (Human); 22778 (Mouse); 305501 (Rat) |
| 应用文献 | |
| 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. Epstein-Barr virus utilizes Ikaros in regulating its latent-lytic switch in B cells. | |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | E. coli-derived recombinant human lkaros Ser427-Ser519 Accession # Q13422 |
| 生物活性 | Human |
| 标记 | Unconjugated |
| 溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 背景 | 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. |
| 运输条件 | Blue Ice |
| 存放说明 | 4℃ |
| 参考文献 |
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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 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. The VPAC promoter was detected by standard PCR. | |
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 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. |
