Human SLC9A3R1 (EBP50/NHERF-1) knockout HCT116 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: 1 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 HCT116 cell line (AB266876)
Lanes 1- 2 : Merged signal (red and green). Green - ab109430 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109430 was shown to react with EBP50/NHERF-1 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266876 (knockout cell lysate ab257281) was used. Wild-type HCT116 and SLC9A3R1 knockout HCT116 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. ab109430 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-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 HCT116 cell lysate at 20 µg
Lane 2:
SLC9A3R1 knockout HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HCT116 cell line (ab266876)
Predicted band size: 38 kDa
Observed band size: 48 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SLC9A3R1 (EBP50/NHERF-1) knockout HCT116 cell line (AB266876)
Homozygous : 1 bp deletion in exon1
Reactivity data
Product details
Recommended control: Human wild-type HCT116 cell line (ab255451). 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
McCoY5a + 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: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB247858
Anti-EBP50/NHERF-1 antibody [EPR5562] - BSA and Azide free
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Primary Antibodies
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Anti-BTBD17 antibody [EPR9593(B)] - BSA and Azide free
RabMAb|Recombinant
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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