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NPC2 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 insertion in exon 1.

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Images

Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749), expandable thumbnail
  • Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749), expandable thumbnail
  • Cell Culture - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749), expandable thumbnail
  • Sanger Sequencing - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749), expandable thumbnail
  • Sanger Sequencing - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

Alternative names

EDDM1, Epididymal protein 1, Epididymal secretory protein, Epididymal secretory protein E1, Human epididymis-specific protein 1, NPC intracellular cholesterol transporter 2, NPC2_HUMAN, Niemann-Pick disease type C2, Niemann-Pick disease type C2 protein, Tissue specific secretory protein, hE1

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NPC2 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 insertion in exon 1.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1
Concentration
Loading...

Properties

Gene name
NPC2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

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

Cell culture

Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female

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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The Niemann Pick C2 protein (NPC2) is a small intracellular cholesterol transport protein weighing approximately 16 kDa. Alternately referred to as "NPC2 protein" or "NPC 293" it binds with lipids and is predominantly expressed in the lysosome. This protein plays a critical role in the intracellular lipid transport process. NPC2 forms part of a family of proteins tasked with moving cholesterol and other related lipids within lysosomal compartments.

Biological function summary

NPC2 protein facilitates the transfer of cholesterol from lysosomes to other parts of the cell. It operates in tandem with its partner protein Niemann Pick C1 (NPC1) to mediate the egress of cholesterol. The NPC2 protein docks to NPC1 within the lysosomal membrane forming an essential complex in lipid transport. This function is pivotal for the maintenance of cellular cholesterol homeostasis influencing various cellular and systemic functions linked to cholesterol metabolism.

Pathways

NPC2 protein participates prominently in the cholesterol efflux pathways. It associates with the NPC1 protein to regulate the movement of cholesterol from the lysosomal membrane to cytoplasmic areas where it can be utilized or stored. This pathway links NPC2 to significant biological processes like the homeostasis of lipoprotein-derived cholesterol influencing the SREBP pathway which governs lipid synthesis and uptake. Such interactions highlight NPC2's involvement in regulating cellular lipid balance.

Associated diseases and disorders

NPC2 protein dysfunction notably links to Niemann-Pick disease type C a lipid storage disorder. Mutations in the genes encoding NPC2 or NPC1 disrupt cholesterol transportation leading to abnormal lipid accumulation and progressive neurodegeneration. Additionally this protein's dysregulation is indirectly connected to atherosclerosis as such disruptions can result in altered cellular cholesterol management highlighting its interaction with proteins involved in cardiovascular diseases.

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