Human FTL knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
FTL KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
FRIL_HUMAN, FTL, Ferritin L chain, Ferritin L subunit, Ferritin light chain, Ferritin light polypeptide, LFTD, NBIA 3, ferritin light polypeptide like 3
- WB
Lab
Western blot - Human FTL knockout HeLa cell line (AB265533)
Lanes 1-2 : Merged signal (red and green). Green - ab109373 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109373 Anti-FTL was shown to specifically react with Ferritin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265533 (knockout cell lysate ab256926) was used. Wild-type and FTL knockout samples were subjected to SDS-PAGE. ab109373 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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-Ferritin Light Chain antibody [EPR5260] (<a href='/en-us/products/primary-antibodies/ferritin-light-chain-antibody-epr5260-ab109373'>ab109373</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Ferritin knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human FTL knockout HeLa cell line (ab265533)
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 20 kDa
Observed band size: 20 kDa
false
- WB
Lab
Western blot - Human FTL knockout HeLa cell line (AB265533)
ab218400 Anti-Ferritin Light Chain antibody [FTL/1386] was shown to specifically react with Ferritin Light Chain in wild-type HeLa cells (ab255928). Loss of signal was observed when knockout cell line ab265534 (knockout cell lysate ab256927) was used. Wild-type and Ferritin Light Chain knockout samples were subjected to SDS-PAGE. ab218400 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Ferritin Light Chain antibody [FTL/1386] (<a href='/en-us/products/primary-antibodies/ferritin-light-chain-antibody-ftl-1386-ab218400'>ab218400</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
FTL knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human FTL knockout HeLa cell line (ab265533)
Secondary
All lanes:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution
Predicted band size: 20 kDa
Observed band size: 20 kDa
false
- Cell Culture
Unknown
Cell Culture - Human FTL knockout HeLa cell line (AB265533)
Representative images of FTL knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human FTL knockout HeLa cell line (AB265533)
Allele-2 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human FTL knockout HeLa cell line (AB265533)
Allele-1 : 1 bp deletion in exon 1.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human FTL knockout HeLa cell line (AB265533)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized FTL KO HeLa (FTL knockout human cervical adenocarcinoma epithelial cell) ab265533 cells labelling Ferritin Light Chain with ab313563 at 1/50 (10.12 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mainly cytoplasmic staining in wildtype HeLa cells and showing no staining in FTL knockout HeLa cells.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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 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
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB125066
Anti-Glutathione Peroxidase 4 antibody [EPNCIR144]
BOND RX™ Validated|KO Validated|RabMAb|Recombinant|Lab Essentials
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (AB125066)](https://content.abcam.com/products/images/glutathione-peroxidase-4-antibody-epncir144-ab125066--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img436422.jpg)
- 5
- 13
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com