Human EIF2AK4 knockout THP1 cell line
- Advanced Validation
- What is this?
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EIF2AK4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
eIF-2-alpha kinase GCN2, GCN2, Eukaryotic translation initiation factor 2-alpha kinase 4, EIF2AK4, GCN2-like protein, KIAA1338
- NGS
Lab
Next Generation Sequencing - Human EIF2AK4 knockout THP1 cell line (AB290213)
21 bp deletion (allele 1), 22 bp deletion (allele 2), 23 bp deletion (allele 3) and 24 bp deletion (allele 4) in exon 6, CCDS42016.1
- WB
Lab
Western blot - Human EIF2AK4 knockout THP1 cell line (AB290213)
Western blot : Anti-GCN2 antibody [EPR5970(2)] ab134053 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 187 kDa in Wild-type THP-1 cell lysates with no signal observed at this size in EIF2AK4 knockout THP-1 cell line. To generate this image, 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-GCN2 antibody [EPR5970(2)] (<a href='/en-us/products/primary-antibodies/gcn2-antibody-epr59702-ab134053'>ab134053</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 cell lysates at 20 µg
Lane 2:
Western blot - Human EIF2AK4 knockout THP1 cell line (ab290213) at 20 µg
Lane 3:
Wild-type HEK-293T ab255449 cell lysates at 20 µg
Lane 4:
Western blot - Human EIF2AK4 (GCN2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-eif2ak4-gcn2-knockout-hek-293t-cell-line-ab267247'>ab267247</a>) at 20 µg
Lane 5:
HT1080 cell lysates at 20 µg
Lane 6:
HepG2 cell lysates at 20 µg
Lane 7:
Ramos cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 187 kDa
Observed band size: 187 kDa
false
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Primary Antibodies
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Primary Antibodies
AB169528
Anti-eIF2A antibody [EPR11042]
20ul selling size|RabMAb|Recombinant|KO Validated
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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 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
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Viability
~ 60%
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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 4x10<sup>5</sup> cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Cultures should be monitored daily.<br>5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x10<sup>5</sup> cells/mL.Small amounts of fresh media can be added until cell number/viability improves.</p>
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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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