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AB280875

人APOE knockout Hep G2 cell line

Human APOE knockout Hep G2 cell line

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(1 Publication)

APOE KO cell line available to order. KO validated. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

查看别名

AD2, APOEA, APOE_HUMAN, Apolipoprotein E, Apolipoprotein E3, Apoprotein, LDLCQ5, LPG

3 Images
Western blot - Human APOE knockout Hep G2 cell line (AB280875)
  • WB

Lab

Western blot - Human APOE knockout Hep G2 cell line (AB280875)

False colour image of Western blot : Anti-Apolipoprotein E antibody [EP1374Y] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab52607 was shown to bind specifically to Apolipoprotein E. A band was observed at 34-37 kDa in wild-type HepG2 cell lysates with no signal observed at this size in APOE knockout cell line. To generate this image wild-type and APOE knockout HepG2 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (<a href='/products/primary-antibodies/apolipoprotein-e-antibody-ep1374y-ab52607'>ab52607</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

APOE knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human APOE knockout Hep G2 cell line (ab280875)

Lane 3:

Human Liver cell lysate at 20 µg

Lane 4:

Human Kidney cell lysate at 20 µg

Predicted band size: 36 kDa

Observed band size: 34 kDa,34-37 kDa

false

Western blot - Human APOE knockout Hep G2 cell line (AB280875)
  • WB

Lab

Western blot - Human APOE knockout Hep G2 cell line (AB280875)

False colour image of Western blot : Anti-Apolipoprotein E antibody [EPR19392] staining at 1/2000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab183597 was shown to bind specifically to Apolipoprotein E. A band was observed at 34 kDa in wild-type HepG2 cell lysates with no signal observed at this size in APOE knockout cell line. To generate this image wild-type and APOE knockout HepG2 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Apolipoprotein E antibody [EPR19392] (<a href='/products/primary-antibodies/apolipoprotein-e-antibody-epr19392-ab183597'>ab183597</a>) at 1/2000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

APOE knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human APOE knockout Hep G2 cell line (ab280875)

Lane 3:

Human Liver cell lysate at 20 µg

Lane 4:

Human Kidney cell lysate at 20 µg

Predicted band size: 36 kDa

Observed band size: 34 kDa

false

Sanger Sequencing - Human APOE knockout Hep G2 cell line (AB280875)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human APOE knockout Hep G2 cell line (AB280875)

关键信息

细胞类型

Hep G2

种属

Human

组织

Liver

形式

Liquid

form

敲除验证

Sanger Sequencing

疾病

Hepatocellular Carcinoma

反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

产品详情

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.

规格

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab280875-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab275467 Human wild-type Hep G2 cell line", "number":"AB280875-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab280875 Human APOE knockout Hep G2 cell line", "number":"AB280875-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab280875-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab280875 Human APOE knockout Hep G2 cell line", "number":"AB280875-CMP01", "productcode":"" } ] } } }

性能和储存信息

基因名称
APOE
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Sanger Sequencing
运输条件
Dry Ice
推荐的短期储存条件
-196°C
推荐的长期储存条件
-196°C

处理步骤

初始处理指南

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.

传代培养指南
  • 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.
培养基

MEM + 10% FBS

低温储藏试剂

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

补充信息

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

Apolipoprotein E (ApoE) also known as apolipoprotein e or apoE is a major protein involved in lipid metabolism. It has an approximate molecular weight of 34 kDa. This protein is mainly produced in the liver and brain where it plays a critical role in transporting lipoproteins fat-soluble vitamins and cholesterol. ApoE exists in three common isoforms: ApoE2 ApoE3 and ApoE4 each having different impacts on lipid binding and metabolic processes. Scientists often use an ApoE ELISA kit to quantify this protein in various samples providing insights into its expression in health and disease.
Biological function summary

ApoE mediates the binding internalization and catabolism of these lipoprotein particles facilitating their interaction with specific cell-surface receptors such as the LDL receptor. This protein operates as part of a complex that includes various other apolipoproteins and lipid molecules. The study of mouse apoe using tools like a mouse apoe ELISA provides valuable data due to its similar physiological functions in lipid transport and metabolism.

Pathways

In the lipid metabolism pathway ApoE interacts with proteins such as the LDL receptor influencing the clearance of chylomicron remnants and VLDL from the bloodstream. In the cardiovascular disease pathway this protein impacts cholesterol levels and promotes plaques stabilization. ApoE's role in these pathways offers insights into its interaction with related proteins like apolipoprotein B and LDL receptor which are critical for maintaining lipid equilibrium.

In Alzheimer's disease ApoE4 isoform has a higher risk factor compared to ApoE3 and ApoE2 contributing to amyloid plaque formation through interactions with amyloid precursor protein. In cardiovascular diseases ApoE abnormalities influence atherosclerosis development with ApoE-deficient models showing increased susceptibility. ApoE's links to these diseases also connect it to other key proteins such as presenilin-1 in Alzheimer's disease and apolipoprotein B in cardiovascular disorders highlighting its extensive biological impact.

细胞培养

生物安全等级

EU: 1 US: 1

贴壁/悬浮

Adherent

性别

Male

产品实验方案

文献 (1)

Recent publications for all applications. Explore the full list and refine your search

Nature microbiology 9:1499-1512 PubMed38548922

2024

Crimean-Congo haemorrhagic fever virus uses LDLR to bind and enter host cells.

Applications

Unspecified application

Species

Unspecified reactive species

Vanessa M Monteil,Shane C Wright,Matheus Dyczynski,Max J Kellner,Sofia Appelberg,Sebastian W Platzer,Ahmed Ibrahim,Hyesoo Kwon,Ioannis Pittarokoilis,Mattia Mirandola,Georg Michlits,Stephanie Devignot,Elizabeth Elder,Samir Abdurahman,Sándor Bereczky,Binnur Bagci,Sonia Youhanna,Teodor Aastrup,Volker M Lauschke,Cristiano Salata,Nazif Elaldi,Friedemann Weber,Nuria Monserrat,David W Hawman,Heinz Feldmann,Moritz Horn,Josef M Penninger,Ali Mirazimi
View all publications

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