FRK Stable Knockout Human H1299 Cell Line
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产品名称: FRK Stable Knockout Human H1299 Cell Line
产品型号: T9505
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简单介绍
FRK Stable Knockout Human H1299 Cell Line
FRK Stable Knockout Human H1299 Cell Line
的详细介绍
|
Print Version |
BioSafety Level |
II |
Organism |
Homo sapiens |
SourceOrgan |
Lung |
Growth Properties |
Adherent |
Morphology |
Polygonal |
Description |
FRK Stable Knockout Human H1299 Cell Line is established via CRISPR/Cas9 mediated frameshift mutation of the FRK gene (NM_002031) using the following guide RNA sequence: GAGGACTTGAGCTTCCGAGC. FRK gene has important functions during G1 and S phase of the cell cycle and thus can be used to study cell proliferation. This cell line is sequenced verified to be biallelic knockout. |
Procedure Overview |
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Clones |
1C10 |
Passage Number |
P4 |
Applications |
For Research Use Only |
Propagation |
Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow Ⅱ medium available in ABM, Cat. No. TM002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO2), 5%.
Selection: 0.5 µg/ml Puromycin (G264) |
Subculturing |
1. Remove and discard culture medium.
2. Add 2.0mL of Trypsin-EDTA(TM050) solution to flask and observe cells under an inverted microscope until cell layer is dispersed. Note: Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
3. Centrifuge cells at 1500rpm for 3 minutes to pellet.
4. Aspirate out trypsin, leaving pellet undisturbed.
5. Resuspend pellet in fresh culture medium and plate in new culture vessel.
6. Incubate cultures at 37°C. |
Preservation |
1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.
2. Storage Temperature: Liquid nitrogen vapour phase. |
Pictures |
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Pictures |
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Pictures |
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Quality Control |
1) 0.5 µg/ml of puromycin (G264) was used for clonal selection; 2) FRK gene knockout confirmed via surveryor assay and sanger sequencing of targeted genomic region |
Disclaimer |
1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers. All sales are final.
2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).
3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.
4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. |