Analysis of Ikaros Expression Left Panel - Flow cytometric analysis of Ikaros expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear (PBMC) cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; dashed line histogram) or Purified Mouse Anti-Ikaros antibody (Cat. No. 564475; solid line histogram). The cells were washed and stained with PE Goat Anti-Mouse Ig (Cat. No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. Middle Panel - Western blot analysis of Ikaros expressed by human PBMC. A PBMC lysate was electrophoresed (SDS-PAGE) and the cellular proteins were transferred to a membrane where they were probed using Purified Mouse Anti-Ikaros antibody at concentrations of 4 (lane 1), 1 (lane 2), and 0.25 μg/ml (lanes 3). Ikaros was identified in bands ranging in size from ~50-70 kDa. Right Panel - Ikaros staining of human tonsil. Tissue sections were prepared from formalin-fixed paraffin-embedded tonsils. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the sections were stained with either Purified Mouse IgG1 κ Isotype Control (Left Tonsil Section, 20x original) or Purified Mouse Anti-Ikaros antibody (Middle and Right Tonsil sections at 20x and 40x original, respectively). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissue was counterstained with Hematoxylin. Ikaros staining is detected in the nuclei of some lymphocytes.
Analysis of Ikaros Expression Left Panel - Flow cytometric analysis of Ikaros expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear (PBMC) cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; dashed line histogram) or Purified Mouse Anti-Ikaros antibody (Cat. No. 564475; solid line histogram). The cells were washed and stained with PE Goat Anti-Mouse Ig (Cat. No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. Middle Panel - Western blot analysis of Ikaros expressed by human PBMC. A PBMC lysate was electrophoresed (SDS-PAGE) and the cellular proteins were transferred to a membrane where they were probed using Purified Mouse Anti-Ikaros antibody at concentrations of 4 (lane 1), 1 (lane 2), and 0.25 μg/ml (lanes 3). Ikaros was identified in bands ranging in size from ~50-70 kDa. Right Panel - Ikaros staining of human tonsil. Tissue sections were prepared from formalin-fixed paraffin-embedded tonsils. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the sections were stained with either Purified Mouse IgG1 κ Isotype Control (Left Tonsil Section, 20x original) or Purified Mouse Anti-Ikaros antibody (Middle and Right Tonsil sections at 20x and 40x original, respectively). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissue was counterstained with Hematoxylin. Ikaros staining is detected in the nuclei of some lymphocytes.
Product Details
BD Pharmingen™
IKZF1; LYF1; hIk-1; IK1; ZNFN1A1
Human (QC Testing), Mouse (Lack of Reactivity Confirmed in Development)
Mouse IgG1, κ
Human Ikaros Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Immunochemistry, Western blot (Tested During Development)
0.5 mg/ml
AB_2738823
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
Retrievagen A (pH 6.0) RUO
Cat No.
550524
Purified Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
554121
Transcription Factor Buffer Set RUO
Size
25 Tests
Cat No.
562725
PE Goat Anti-Mouse Ig (Multiple Adsorption) RUO
Size
0.2 mg
Cat No.
550589
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564475 Rev. 1
Antibody Details
R32-1149
The R32-1149 monoclonal antibody specifically binds to Ikaros, which is encoded by IKZF1 (IKAROS family zinc finger 1). Ikaros is also known as Lymphoid transcription factor LyF-1 (LYF1) and ZNFN1A1 [Zinc finger protein, subfamily 1A, 1 (Ikaros)]. Ikaros belongs to the IKAROS family of zinc-finger transcription factors. This DNA-binding transcription factor forms homodimers and heterodimers with other IKAROS family members. It is involved in the development of lymphoid tissues and remains abundantly expressed in the thymus and by cells within peripheral lymphoid tissues. Ikaros functions in the maturation, differentiation, and homeostasis of various hematopoietic cells including T lymphocytes, B cells, NK cells, and neutrophils. Ikaros regulates the expression of a number of genes including various genes expressed during early stages of B- and T-cell development. Several alternatively-spliced human Ikaros isoforms have been described. Abnormal expression of different Ikaros isoforms has been associated with hematologic malignancies. Flow cytometric analysis of 293F cells transfected with members of the IKAROS family has demonstrated that clone R32-1149 is negative on Helios, Aiolos, and Eos transfectants.
564475 Rev. 1
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
564475 Rev.1
Citations & References
Development References (5)
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Georgopoulos K. Haematopoietic cell-fate decisions, chromatin regulation and ikaros. Nat Rev Immunol. 2002; 2(3):162-174. (Biology). View Reference
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Molnar A, Wu P, Largespada DA, et al. The Ikaros gene encodes a family of lymphocyte-restricted zinc finger DNA binding proteins, highly conserved in human and mouse. J Immunol. 1996; 156(2):585-592. (Biology). View Reference
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Nietfeld W, Meyerhans A. Cloning and sequencing of hIk-1, a cDNA encoding a human homologue of mouse Ikaros/LyF-1. Immunol Lett. 1996; 49(1-2):139-141. (Biology). View Reference
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Westman BJ, Mackay JP, Gell D. Ikaros: a key regulator of haematopoiesis. Int J Biochem Cell Biol. 2002; 34(10):1304-1307. (Biology). View Reference
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Yoshida T, Landhuis E, Dose M, et al. Transcriptional regulation of the Ikzf1 locus. Blood. 2013; 122(18):3149-3159. (Biology). View Reference
564475 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
