IKZF1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 2 bp insertion, 1 bp insertion; Frameshift: 100%.
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Key facts
- Cell type
Jurkat
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Mutation description
Knockout achieved by CRISPR/Cas9; X = 2 bp insertion, 1 bp insertion; Frameshift: 100%
Alternative names
CLL associated antigen KW 6, DNA-binding protein Ikaros, Hs.54452, IK1, IKAROS, IKAROS family zinc finger 1 (Ikaros), IKZF1_HUMAN, Ikaros (zinc finger protein), Ikaros family zinc finger protein 1, LYF1, Lymphoid transcription factor LyF-1, PRO0758, ZNFN1A1, Zinc finger protein subfamily 1A 1, Zinc finger protein subfamily 1A 1 (Ikaros), Zinc finger protein, subfamily 1A, member 1, hIk 1
Recommended products
IKZF1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 2 bp insertion, 1 bp insertion; Frameshift: 100%.
Key facts
- Cell type
Jurkat
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Mutation description
Knockout achieved by CRISPR/Cas9; X = 2 bp insertion, 1 bp insertion; Frameshift: 100%
- Disease
Non-Hodgkin Lymphoma
- Concentration
Properties
- Gene name
IKZF1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Next Generation Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 1 US: 1
- Adherent/suspension
Suspension
- Gender
Male
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water for bath approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
- Do not allow cell density to exceed 3x106 cells/mL.
- Culture medium
RPMI + 10% FBS
- Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type Jurkat cell line (Human wild-type Jurkat cell line ab271143). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Ikaros also known as IKZF1 is a transcription factor. You also find it being referred to as the Ikaros protein or Ikaros code. It has a molecular mass of about 58-60 kDa. Ikaros expresses mostly in hematopoietic cells including precursors from which various blood cell types derive. It plays a significant role in regulating gene expression and cellular differentiation modulating the chromatin structure to control access to DNA.
- Biological function summary
Ikaros protein functions in the regulation of lymphoid cell development. It is a part of a chromatin-remodeling complex influencing chromatin accessibility. Through its zinc finger domains it binds to specific DNA sequences controlling genes necessary for lymphocyte differentiation and proliferation. Ikaros influences various cell types within the immune system especially affecting T and B cell development and also regulates cytokine receptor genes necessary for proper immune function.
- Pathways
The Ikaros protein is critical to lymphocyte signaling pathways particularly the JAK/STAT and NF-κB pathways. In these pathways Ikaros interacts with other transcription factors like GATA3 and STAT5 integrating signals that are important for immune cell function and development. This integration allows a coordinated response to extracellular signals ensuring effective immune responses or tolerance.
- Associated diseases and disorders
Ikaros is associated with leukemia and lymphomas. Alterations or mutations in ikaros gene can lead to misregulation of its target genes contributing to the development of these diseases by disturbing normal hematopoietic processes. Moreover its interaction with TAL1 protein is significant in certain leukemias where disturbances in the balance of their activities can lead to malignancies. Understanding Ikaros’s role provides insights for therapeutic approaches in these blood disorders.
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3 product images
Next Generation Sequencing - Human IKZF1 knockout Jurkat cell line (ab274920)
Knockout achieved by CRISPR/Cas9; X = 2 bp insertion, 1 bp insertion; Frameshift: 100%
Western blot - Human IKZF1 knockout Jurkat cell line (ab274920)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Ikaros antibody [EPR13790] ab191394 observed at 50-70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-Ikaros antibody [EPR13790] ab191394 was shown to react with Ikaros in wild-type Jurkat cells in western blot with loss of signal observed in IKZF1 knockout cell line ab274920 (knockout cell lysate Human IKZF1 knockout Jurkat cell lysate ab274978). Wild-type and IKZF1 knockout Jurkat cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-Ikaros antibody [EPR13790] ab191394 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Ikaros antibody [EPR13790] (Anti-Ikaros antibody [EPR13790] ab191394) at 1/10000 dilution
Lane 1: Wild-type Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 2: IKZF1 knockout Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 3: Daudi (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 50-70 kDa
Next Generation Sequencing - Human IKZF1 knockout Jurkat cell line (ab274920)
2 bp deletion (allele 1) and 1 bp deletion (allele 2) after Arg50 of the WT protein
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