Human JAK1 knockout HCT116 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human JAK1 knockout HCT116 cell line (AB286460)
Western blot : Anti-JAK1 antibody [EPR349(N)] (ab133666) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133666 was shown to bind specifically to JAK1. A band was observed at 120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in JAK1 knockout cell line. To generate this image, wild-type and JAK1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-JAK1 antibody [EPR349(N)] (<a href='/en-us/products/primary-antibodies/jak1-antibody-epr349n-ab133666'>ab133666</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human JAK1 knockout HCT116 cell line (ab286460)
Lane 2:
JAK1 knockout HCT 116 cell lysate at 20 µg
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 120 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human JAK1 knockout HCT116 cell line (AB286460)
247 bp deletion after Leu234
Complementary Products
Primary Antibodies
AB225636
Anti-HLA Class 1 ABC antibody [EPR22172]
BOND RX™ Validated|RabMAb|Recombinant|20ul selling size
![Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (AB225636)](https://content.abcam.com/products/images/hla-class-1-abc-antibody-epr22172-ab225636--flow-cytometry-img27275.jpg)
- 4
- 1
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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 bath for 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 seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. 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 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
McCoY5a + 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
This enzyme participates significantly in the immune response and hematopoiesis by being part of the JAK-STAT signaling pathway complex. When activated JAK1 phosphorylates specific tyrosine residues on the receptor creating docking sites for STAT proteins which then get phosphorylated by JAK1. This phosphorylation allows STAT proteins to dimerize and translocate to the nucleus affecting gene expression involved in growth survival and differentiation processes.
Pathways
JAK1 is importantly engaged in the JAK-STAT pathway and the Interferon signaling pathway. Within these pathways JAK1 collaborates closely with other proteins like JAK2 TYK2 and STAT proteins. It acts as a mediator of the cellular response to a variety of cytokines and growth factors playing a role in the regulation of immune responses and cell proliferation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
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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