Human FHL1 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human FHL1 knockout HeLa cell line (AB266011)
Lanes 1-4 : Merged signal (red and green). Green - ab255828 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.
ab255828 Anti-FHL1 antibody [EPR22842-95] was shown to specifically react with FHL1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266011 (knockout cell lysate ab257952) was used. Wild-type and FHL1 knockout samples were subjected to SDS-PAGE. ab255828 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-FHL1 antibody [EPR22842-95] (<a href='/en-us/products/primary-antibodies/fhl1-antibody-epr22842-95-ab255828'>ab255828</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
FHL1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human FHL1 knockout HeLa cell line (ab266011)
Lane 3:
HEK293T cell lysate at 20 µg
Lane 4:
Daudi 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: 36 kDa
Observed band size: 32 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human FHL1 knockout HeLa cell line (AB266011)
Allele-1 : 1 bp deletion in exon 4.
- Sanger seq
Lab
Sanger Sequencing - Human FHL1 knockout HeLa cell line (AB266011)
Allele-2 : Insertion of the selection cassette in exon 4.
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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, 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
FHL1 integrates into diverse cellular mechanisms by engaging in protein-protein interactions. It acts as part of complexes that regulate transcription cell differentiation and development. The involvement of FHL1 in muscle growth and maintenance is significant partly due to its expression pattern. In skeletal muscles it helps adapt muscle response to mechanical stress while in cardiac tissue it participates in muscle contractility and integrity.
Pathways
FHL1 plays important roles in both the muscle differentiation pathway and the MAPK/ERK signaling pathway. It influences the transcriptional regulation essential for muscle cell growth and differentiation. FHL1 interacts with proteins such as MyoD during muscle formation and MEF2 in the MAPK/ERK pathway modulating cellular responses to external signals and stressors.
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
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