Human Ikaros Antibody
R&D Systems, part of Bio-Techne | Catalog # AF4984
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Chromatin Immunoprecipitation (ChIP), CyTOF-ready, Immunocytochemistry, Intracellular Staining by Flow Cytometry, Simple Western, Western Blot
Cited:
Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Specifications
Immunogen
E. coli-derived recombinant human lkaros
Ser427-Ser519
Accession # Q13422
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 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. TheVPACpromoter 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.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
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
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
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
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
Sample: Nalm-6 human Pre-B acute lymphocytic leukemia cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 1 review rated 4 using AF4984 in the following applications:
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.
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
Gene Symbol
IKZF1
UniProt
Additional Ikaros Products
Product Documents for Human Ikaros Antibody
Product Specific Notices for Human Ikaros Antibody
For research use only
Loading...
Loading...
Loading...
Loading...
Loading...