Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line
- Advanced Validation
- What is this?
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NHERF1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 1.
View Alternative Names
EBP 50, Ezrin-radixin-moesin-binding phosphoprotein 50, NHERF, NHERF-1, NHRF1_HUMAN, NPHLOP2, Na(+)/H(+) exchange regulatory cofactor NHE RF, Na(+)/H(+) exchange regulatory cofactor NHE-RF1, Na+/H+ exchange regulatory co factor, Regulatory cofactor of Na(+)/H(+) exchanger, Slc9a3r1, Sodium-hydrogen exchanger regulatory factor 1, Sodium/hydrogen exchanger regulatory factor 1, Solute carrier family 9 (sodium/hydrogen exchanger) member 3 regulator 1, Solute carrier family 9 (sodium/hydrogen exchanger), isoform 3 regulatory factor 1, Solute carrier family 9 isoform 3 regulatory factor 1, Solute carrier family 9 isoform A3 regulatory factor 1, Solute carrier family 9 member 3 regulator 1
- WB
Lab
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (AB264914)
False colour image of Western blot : Anti-EBP50/NHERF-1 antibody [EPR5562] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab109430 was shown to bind specifically to EBP50/NHERF-1. A band was observed at 46 kDa in wild-type HeLa cell lysates with no signal observed at this size in SLC9A3R1 knockout cell line ab264914 (knockout cell lysate ab257280). To generate this image wild-type and SLC9A3R1 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (<a href='/en-us/products/primary-antibodies/ebp50-nherf-1-antibody-epr5562-ab109430'>ab109430</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SLC9A3R1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (ab264914)
Lane 2:
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-slc9a3r1-ebp50-nherf-1-knockout-hela-cell-lysate-ab257280'>ab257280</a>)
Predicted band size: 38 kDa
Observed band size: 46 kDa
false
- WB
Lab
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (AB264914)
False colour image of Western blot : Anti-EBP50/NHERF-1 antibody staining at 1 μg/ml shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab88238 was shown to bind specifically to EBP50/NHERF-1. A band was observed at 46 kDa in wild-type HeLa cell lysates with no signal observed at this size in SLC9A3R1 knockout cell line ab264914 (knockout cell lysate ab257280). To generate this image wild-type and SLC9A3R1 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-EBP50/NHERF-1 antibody (<a href='/en-us/products/primary-antibodies/ebp50-nherf-1-antibody-ab88238'>ab88238</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SLC9A3R1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (ab264914)
Lane 2:
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-slc9a3r1-ebp50-nherf-1-knockout-hela-cell-lysate-ab257280'>ab257280</a>)
Predicted band size: 38 kDa
Observed band size: 46 kDa
false
- Cell Culture
Unknown
Cell Culture - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (AB264914)
Representative images of SLC9A3R1 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 SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (AB264914)
Homozygous : 19 bp deletion in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
EBP50 regulates cellular functions by organizing signaling complexes at the plasma membrane. It is involved in processes such as ion transport signal transduction and cell proliferation. EBP50 often acts as part of a multiprotein complex contributing to the stabilization and regulation of membrane receptors and channels. By regulating these components EBP50 affects a variety of cellular responses to external stimuli.
Pathways
EBP50 plays important roles in cellular signaling pathways like the PTH (parathyroid hormone) receptor signaling and CFTR (cystic fibrosis transmembrane conductance regulator) channel regulation. It interacts with proteins such as ezrin and PLCβ (phospholipase C beta) which facilitates the signaling cascades critical for maintaining proper cell function. By modulating these pathways EBP50 has a significant impact on maintaining cellular homeostasis and responding to changing physiological conditions.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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Primary Antibodies
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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