Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749)
Lanes 1-3 : Merged signal (red and green). Green - ab218192 observed at 16-18 kDa. Red - loading control ab8245 observed at 36 kDa.
ab218192 Anti-Niemann Pick C2 antibody [EPR19993-145-1] was shown to specifically react with Niemann Pick C2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266749 (knockout cell lysate ab258079) was used. Wild-type and Niemann Pick C2 knockout samples were subjected to SDS-PAGE. ab218192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 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-Niemann Pick C2 antibody [EPR19993-145-1] (<a href='/en-us/products/primary-antibodies/niemann-pick-c2-antibody-epr19993-145-1-ab218192'>ab218192</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (ab266749)
Lane 3:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa
false
- WB
Lab
Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749)
Lanes 1-3 : Merged signal (red and green). Green - ab207158 observed at 16-18 kDa. Red - loading control ab8245 observed at 36 kDa.
ab207158 Anti-Niemann Pick C2 antibody [EPR19993] was shown to specifically react with Niemann Pick C2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266749 (knockout cell lysate ab258079) was used. Wild-type and Niemann Pick C2 knockout samples were subjected to SDS-PAGE. ab207158 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 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-Niemann Pick C2 antibody [EPR19993] (<a href='/en-us/products/primary-antibodies/niemann-pick-c2-antibody-epr19993-ab207158'>ab207158</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (ab266749)
Lane 3:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa
false
- Cell Culture
Unknown
Cell Culture - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749)
Representative images of NPC2 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T 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 M5000 microscope.
- Sanger seq
Lab
Sanger Sequencing - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749)
Sequencing chromatogram displaying sequence edit in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749)
Homozygous : 1 bp insertion in exon 1
Complementary Products
Primary Antibodies
AB218192
Anti-Niemann Pick C2 antibody [EPR19993-145-1]
RabMAb|Recombinant|KO Validated
![Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (AB218192)](https://content.abcam.com/products/images/niemann-pick-c2-antibody-epr19993-145-1-ab218192--flow-cytometry-intracellular-img2670.jpg)
- 5
- 1
Reactivity data
Product details
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?
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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