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AB258079

Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate

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NPC2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

View 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

3 Images
Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (AB258079)
  • WB

Lab

Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (AB258079)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)

ab218192 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 4oC 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 (<a href='/en-us/products/cell-lines/human-npc2-niemann-pick-c2-knockout-hek-293t-cell-line-ab266749'>ab266749</a>)

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

Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (AB258079)
  • WB

Lab

Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (AB258079)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)

ab207158 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 4oC 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 (<a href='/en-us/products/cell-lines/human-npc2-niemann-pick-c2-knockout-hek-293t-cell-line-ab266749'>ab266749</a>)

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

Sanger Sequencing - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (AB258079)
  • Sanger seq

Unknown

Sanger Sequencing - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell lysate (AB258079)

Homozygous : 1 bp insertion in exon 1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
NPC2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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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)

  • 5
  • 1
Cell Lines & Lysates

AB266749

Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line

Advanced Validation

Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (AB266749)

  • 0
  • 0
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Primary Antibodies

AB154356

Anti-Lysosomal acid lipase/LAL antibody

Western blot - Anti-Lysosomal acid lipase/LAL antibody (AB154356)

  • 0
  • 0

Product promise

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For full details, please see our Terms & Conditions

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