Human ELK1 knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human ELK1 knockout HeLa cell line (AB261764)
Lanes 1- 2 : Merged signal (red and green). Green - ab188316 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab188316 was shown to react with ELK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261764 (knockout cell lysate ab256904) was used. Wild-type HeLa and ELK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab188316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-ELK1 antibody [EPR1913(2)] (<a href='/en-us/products/primary-antibodies/elk1-antibody-epr19132-ab188316'>ab188316</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ELK1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ELK1 knockout HeLa cell line (ab261764)
Predicted band size: 45 kDa
Observed band size: 55 kDa
false
- WB
Lab
Western blot - Human ELK1 knockout HeLa cell line (AB261764)
Lanes 1- 2 : Merged signal (red and green). Green - ab32106 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32106 was shown to react with ELK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261764 (knockout cell lysate ab256904) was used. Wild-type HeLa and ELK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32106 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 500 dilution and a 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-ELK1 antibody [E277] (<a href='/en-us/products/primary-antibodies/elk1-antibody-e277-ab32106'>ab32106</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ELK1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ELK1 knockout HeLa cell line (ab261764)
Predicted band size: 45 kDa
Observed band size: 55 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ELK1 knockout HeLa cell line (AB261764)
Homozygous : 222 bp insertion in exon 2.
Complementary Products
Primary Antibodies
AB32106
RabMAb|Recombinant|KO Validated|Advanced Validation
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ELK1 antibody [E277] (AB32106)](https://content.abcam.com/products/images/elk1-antibody-e277-ab32106--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img44492.jpg)
- 0
- 0
Primary Antibodies
AB179483
Anti-HIF-1 alpha antibody [EPR16897]
RabMAb|Advanced Validation|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] (AB179483)](https://content.abcam.com/products/images/hif-1-alpha-antibody-epr16897-ab179483--immunocytochemistry-immunofluorescence-img85620.jpg)
- 4
- 9
Cell Lines & Lysates
AB255444
Human VDAC1 (Porin) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
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
Zygosity
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
ELK1 functions in the regulation of gene expression related to cellular proliferation and differentiation. It often forms a ternary complex with serum response factor (SRF) on serum response elements (SREs) to modulate the transcriptional responses of genes important for cell growth. The activation of ELK1 is phosphorylation-dependent primarily by mitogen-activated protein kinases (MAPKs) which further facilitates its role as a transcription regulator. This phosphorylation increases the transcriptional activity enabling ELK1 to actively participate in cellular responses to external signals.
Pathways
ELK1 participates prominently in the MAPK/ERK signaling pathway and the JNK signaling pathway. Through these pathways ELK1 gets activated influencing the transcription of immediate early genes responsible for cell cycle progression and apoptosis. ELK1 interacts with other proteins in these pathways such as MEK and ERK leading to a coordinated response to growth signals. These interactions underline its importance in signal transduction processes and cellular response mechanisms.
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