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AB266258

Human TMEM147 knockout HEK-293T cell line

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TMEM147 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2.

View Alternative Names

NIFIE 14, Protein NIFIE 14, Seven transmembrane domain protein, TM147_HUMAN, Transmembrane protein 147

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Sanger Sequencing - Human TMEM147 knockout HEK-293T cell line (AB266258)
  • Sanger seq

Unknown

Sanger Sequencing - Human TMEM147 knockout HEK-293T cell line (AB266258)

Homozygous : Insertion of the selection cassette in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2

Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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, 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?

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Properties and storage information

Gene name
TMEM147
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.

TMEM147 also known as NBR1 is a transmembrane protein with a molecular mass of approximately 28 kDa. It resides primarily in the endoplasmic reticulum and is widely expressed across various tissues such as the brain and liver. The protein spans the membrane multiple times indicating its role in maintaining cellular structure and function within the membrane environment. TMEM147 is often associated with regulatory processes that affect membrane-related activities.
Biological function summary

TMEM147 interacts with proteins in the ribosome maturation process and is involved in assembling the nuclear pore complex. It forms part of the N-oligosaccharyltransferase (OST) complex which plays an essential role in protein modification through glycosylation. This complex activity helps in the proper folding and stability of proteins necessary for cell function. Therefore TMEM147 influences cellular processes by regulating the maturation and transport of proteins within cells.

Pathways

TMEM147 contributes significantly to the N-glycosylation and protein biosynthesis pathways. Within these pathways the protein interacts with enzymes like STT3A and STT3B which are key catalytic components of the oligosaccharyltransferase complex. Through this interaction TMEM147 ensures proper protein folding and efficiency in glycoprotein processing. It also influences cellular homeostasis and stress responses by coordinating with proteins related to ER stress pathways.

TMEM147 shows a connection to neurodegenerative diseases and certain types of cancer. Studies highlight its involvement in Alzheimer's disease where it interacts with the γ-secretase complex including proteins like presenilin-1 (PSEN1) affecting amyloid-beta precursor protein processing. In cancer abnormal TMEM147 expression correlates with altered N-glycosylation patterns leading to changes in tumor cell behavior. This association suggests TMEM147's potential role as a biomarker or therapeutic target in disease management.

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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