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AB266095

人VKORC1 knockout HEK-293T cell line

Human VKORC1 knockout HEK-293T cell line

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VKORC1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 20 bp deletion in exon 1. 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

VKORC1, MSTP576, Vitamin K epoxide reductase complex subunit 1, MSTP134, VKOR, UNQ308/PRO351

3 Images
Sanger Sequencing - Human VKORC1 knockout HEK-293T cell line (AB266095)
  • Sanger seq

Unknown

Sanger Sequencing - Human VKORC1 knockout HEK-293T cell line (AB266095)

Allele-2 : 1 bp insertion in exon 1.

Sanger Sequencing - Human VKORC1 knockout HEK-293T cell line (AB266095)
  • Sanger seq

Unknown

Sanger Sequencing - Human VKORC1 knockout HEK-293T cell line (AB266095)

Allele-1 : 20 bp deletion in exon1

Western blot - Human VKORC1 knockout HEK-293T cell line (AB266095)
  • WB

Lab

Western blot - Human VKORC1 knockout HEK-293T cell line (AB266095)

Western blot : Anti-VKORC1 antibody [EPR20245] ab206656 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 15 kDa in Wild-type HEK-293T cell lysates with no signal observed at this size in VKORC1 knockout HEK-293T cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-VKORC1 antibody [EPR20245] (<a href='/products/primary-antibodies/vkorc1-antibody-epr20245-ab206656'>ab206656</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysates at 20 µg

Lane 2:

Western blot - Human VKORC1 knockout HEK-293T cell line (ab266095) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

MCF7 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 18 kDa

Observed band size: 15 kDa

false

关键信息

细胞类型

HEK-293T

种属

Human

组织

Kidney

形式

Liquid

form

敲除验证

Sanger Sequencing,Western blot

突变描述

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 20 bp deletion in exon 1

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, 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.

规格

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性能和储存信息

基因名称
VKORC1
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Sanger Sequencing, Western blot
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

补充信息

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

Vitamin K epoxide reductase complex subunit 1 (VKORC1) also known as VKOR is an enzyme that plays an important role in the vitamin K cycle. This enzyme has a molecular mass of approximately 18 kDa. VKORC1 helps in reducing vitamin K 23-epoxide to its active hydroquinone form which is necessary for the gamma-carboxylation of vitamin K-dependent proteins. These proteins are mainly responsible for blood coagulation. VKORC1 expression is high in the liver where much of the synthesis of clotting factors occurs.
Biological function summary

VKORC1 is integral to the vitamin K cycle an essential process for activating vitamin K-dependent clotting factors. It operates within a complex with its protein partner VKORC1-like 1 (VKORC1L1). This partnership is important for maintaining effective blood coagulation. The active form of vitamin K produced by VKORC1 ensures the activity of several blood clotting factors especially factors II VII IX and X. Without proper function of VKORC1 these factors would remain inactive.

Pathways

The vitamin K cycle is where VKORC1 makes a significant impact and this cycle is vital for blood coagulation processes. Specifically VKORC1 reduces vitamin K epoxide working closely with gamma-glutamyl carboxylase (GGCX) another enzyme that modifies the glutamic acid residues on clotting factors. This modification is important for their proper functioning within the coagulation cascade. The interplay between VKORC1 and GGCX directly connects to the synthesis of active blood coagulation proteins.

VKORC1 mutations are notably linked to warfarin resistance and susceptibility to bleeding disorders. Warfarin is an anticoagulant that targets VKORC1 to prevent clot formation and variations in this protein can lead to altered drug response. Furthermore any dysfunction in VKORC1 may result in vitamin K epoxide reductase deficiency affecting coagulation and leading to increased bleeding risk. Mutations could also affect partner proteins like coagulation factors II and VII leading to disorders related to abnormal blood clotting processes.

质量控制

STR 分析

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

细胞培养

生物安全等级

EU: 2 US: 2

贴壁/悬浮

Adherent

性别

Female

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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

DMEM (High Glucose) + 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.

产品实验方案

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