Human TFG (TRK fused gene) knockout HeLa cell line
- Advanced Validation
- What is this?
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TFG KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and Insertion of the selection cassette in exon 2. 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
FLJ36137, HMSNP, OTTHUMP00000214045, OTTHUMP00000214046, OTTHUMP00000214047, OTTHUMP00000214048, Protein TFG, SPG57, TF6, TFG_HUMAN, TRK fused, TRK fused gene, TRK-fused gene protein, TRKT3, TRKT3 oncogene
- WB
Lab
Western blot - Human TFG (TRK fused gene) knockout HeLa cell line (AB265841)
Lanes 1-4 : Merged signal (red and green). Green - ab150428 observed at 57 kDa. Red - loading control ab8245 observed at 36 kDa.
ab150428 Anti-TRK fused gene antibody [EPR8765(2)(B)] was shown to specifically react with TFG wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265841 (knockout cell lysate ab257738) was used. Wild-type and TFG knockout samples were subjected to SDS-PAGE. ab150428 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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-TRK fused gene antibody [EPR8765(2)(B)] (<a href='/en-us/products/primary-antibodies/trk-fused-gene-antibody-epr87652b-ab150428'>ab150428</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
TFG knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human TFG (TRK fused gene) knockout HeLa cell line (ab265841)
Lane 3:
HAP-1 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
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 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 57 kDa
false
- WB
Unknown
Western blot - Human TFG (TRK fused gene) knockout HeLa cell line (AB265841)
Lanes 1-4 : Merged signal (red and green). Green - ab156866 observed at 57 kDa. Red - loading control ab8245 observed at 36 kDa.
ab156866 Anti-TRK fused gene antibody [EPR8766] was shown to specifically react with TFG in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265841 (knockout cell lysate ab257738) was used. Wild-type and TFG knockout samples were subjected to SDS-PAGE. ab156866 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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-TRK fused gene antibody [EPR8766] (<a href='/en-us/products/primary-antibodies/trk-fused-gene-antibody-epr8766-ab156866'>ab156866</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
TFG knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human TFG (TRK fused gene) knockout HeLa cell line (ab265841)
Lane 3:
HAP-1 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
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 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 57 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TFG (TRK fused gene) knockout HeLa cell line (AB265841)
Allele-1 : 19 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human TFG (TRK fused gene) knockout HeLa cell line (AB265841)
Allele-2 : Insertion of the selection cassette in exon 2.
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Neurotrophin signaling pathways involve TRK proteins facilitating neuronal differentiation survival and plasticity. TRK proteins often form part of larger complexes engaged in transmitting signals from the extracellular environment to intracellular pathways. The TRK proteins interact with neurotrophins like NGF BDNF and NT-3 playing essential roles in nervous system function and development. The interaction between TRK receptors and their ligands activates downstream signaling cascades.
Pathways
TRK proteins significantly contribute to the MAPK/ERK and PI3K/Akt pathways. These pathways control cell growth differentiation and survival. Within these pathways TRK proteins interact with partners such as Ras and Akt coordinating a broad range of cellular processes. They ensure proper cellular responses to external stimuli and maintain homeostasis within the cell.
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
AB156866
Anti-TRK fused gene antibody [EPR8766]
RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-TRK fused gene antibody [EPR8766] (AB156866)](https://content.abcam.com/products/images/trk-fused-gene-antibody-epr8766-ab156866--immunocytochemistry-immunofluorescence-img64252.jpg)
- 0
- 0
Cell Lines & Lysates
AB265788
Human PPID (Cyclophilin 40) knockout HeLa cell line
Advanced Validation
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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