Human IKZF1 knockout Jurkat cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human IKZF1 knockout Jurkat cell line (AB274920)
Lanes 1 - 4 : Merged signal (red and green). Green - ab191394 observed at 50-70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
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 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 ab191394 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Ikaros antibody [EPR13790] (<a href='/en-us/products/primary-antibodies/ikaros-antibody-epr13790-ab191394'>ab191394</a>) 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 2:
Western blot - Human IKZF1 knockout Jurkat cell line (ab274920)
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
Predicted band size: 57 kDa
Observed band size: 50-70 kDa
false
- NGS
Supplier Data
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
- NGS
Supplier Data
Next Generation Sequencing - Human IKZF1 knockout Jurkat cell line (AB274920)
Knockout achieved by CRISPR/Cas9; X = 2 bp insertion, 1 bp insertion; Frameshift : 100%
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (AB191394)](https://content.abcam.com/products/images/ikaros-antibody-epr13790-ab191394--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img110578.jpg)
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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