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AB265346

Human SCNN1D knockout HeLa cell line

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SCNN1D KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 8 and 2 bp deletion in exon 8.

View Alternative Names

Amiloride-sensitive sodium channel subunit delta, DNACH, Delta-ENaC, Delta-NaCH, ENaC Delta, ENaCD, Epithelial Na(+) channel subunit delta, MGC149710, MGC149711, Nonvoltage-gated sodium channel 1 subunit delta, SCNED, SCNND_HUMAN, Sodium channel, nonvoltage gated 1, delta

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Sanger Sequencing - Human SCNN1D knockout HeLa cell line (AB265346)
  • Sanger seq

Unknown

Sanger Sequencing - Human SCNN1D knockout HeLa cell line (AB265346)

Allele-2 : 1 bp deletion in exon 8.

Sanger Sequencing - Human SCNN1D knockout HeLa cell line (AB265346)
  • Sanger seq

Unknown

Sanger Sequencing - Human SCNN1D knockout HeLa cell line (AB265346)

Allele-1 : 2 bp deletion in exon 8.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 8 and 2 bp deletion in exon 8

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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?

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Properties and storage information

Gene name
SCNN1D
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

The SCNN1D gene encodes the delta subunit of the epithelial sodium channel often referred to as ENaC-delta. This subunit forms a part of the multisubunit complex that regulates sodium balance at the epithelial surface. It resides on the cell membrane and plays a mechanical role in facilitating sodium ion transport across epithelial cells. SCNN1D is about 74 kDa in size and is expressed predominantly in renal epithelium and respiratory tissues contributing to sodium homeostasis and fluid balance.
Biological function summary

SCNN1D plays an important role in sodium reabsorption as part of the epithelial sodium channel complex which includes alpha beta and gamma subunits. Together these subunits form a channel responsible for the movement of sodium ions into the cells critical for maintaining electrolyte balance and blood pressure. Its specific contribution enhances the channel's sensitivity and responsiveness to hormonal regulation particularly by aldosterone which modulates electrolyte transport further influencing fluid accumulations in tissues.

Pathways

SCNN1D acts within the aldosterone-regulated sodium reabsorption pathway and the renin-angiotensin-aldosterone system (RAAS). These pathways are essential in regulating blood pressure and fluid balance. SCNN1D works closely with other proteins like the mineralocorticoid receptor which influences ENaC activity by regulating its transcription under the stimulus of aldosterone ensuring facilitating rapid sodium uptake in epithelial cells.

SCNN1D involvement becomes significant in hypertension and cystic fibrosis. Mutations or dysregulation of SCNN1D or its associated pathways can lead to altered blood pressure control due to improper sodium retention. Moreover its role in sodium reabsorption associates this subunit with cystic fibrosis where the balance of ions and fluids in the lungs impacts disease severity. The protein is also linked with cystic fibrosis transmembrane conductance regulator (CFTR) which co-regulates ion transport and channel activity across epithelial surfaces.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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