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AB267267

Human TXNRD2 knockout HEK-293T cell line

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TXNRD2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion 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'.

View Alternative Names

SELZ, Selenoprotein Z, TR, TR 3, TR-beta, TRXR2_HUMAN, Thioredoxin reductase 2, Thioredoxin reductase 2 mitochondrial, Thioredoxin reductase 3, Thioredoxin reductase TR3, Thioredoxin reductase beta, mitochondrial

3 Images
Western blot - Human TXNRD2 knockout HEK-293T cell line (AB267267)
  • WB

Lab

Western blot - Human TXNRD2 knockout HEK-293T cell line (AB267267)

Lanes 1-4 : Merged signal (red and green). Green - ab180493 observed at 57 kDa. Red - loading control ab8245 observed at 36 kDa.

ab180493 Anti-TXNRD2 antibody [EPR12480] was shown to specifically react with TXNRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267267 (knockout cell lysate ab258259) was used. Wild-type and TXNRD2 knockout samples were subjected to SDS-PAGE. ab180493 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-TXNRD2 antibody [EPR12480] (<a href='/en-us/products/primary-antibodies/txnrd2-antibody-epr12480-ab180493'>ab180493</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

TXNRD2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human TXNRD2 knockout HEK-293T cell line (ab267267)

Lane 3:

LNCap cell lysate at 20 µg

Lane 4:

A431 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: 57 kDa

Observed band size: 57 kDa

false

Sanger Sequencing - Human TXNRD2 knockout HEK-293T cell line (AB267267)
  • Sanger seq

Unknown

Sanger Sequencing - Human TXNRD2 knockout HEK-293T cell line (AB267267)

Homozygous : 1 bp insertion in exon1

Cell Culture - Human TXNRD2 knockout HEK-293T cell line (AB267267)
  • Cell Culture

Unknown

Cell Culture - Human TXNRD2 knockout HEK-293T cell line (AB267267)

Representative images of TXNRD2 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 an EVOS M5000 microscope.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

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

Reactivity data

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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?

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Properties and storage information

Gene name
TXNRD2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Thioredoxin reductase 2 (TXNRD2) also known as TRXR2 is an important enzyme in the thioredoxin system. It has a mass of approximately 57 kDa and resides mainly in the mitochondria. TXNRD2 acts as a homodimer and contains a selenocysteine residue at its active site which is essential for its enzymatic activity. TXNRD2 is expressed ubiquitously but shows higher levels in tissues with high metabolic activity such as the heart muscle and brain. Its primary mechanical role involves the reduction of thioredoxin which in turn assists in reducing other proteins by cysteine thiol-disulfide exchange.
Biological function summary

TXNRD2 plays an important role in maintaining cellular redox balance and combating oxidative stress. It is part of the thioredoxin system which together with thioredoxin and thioredoxin peroxidase constitutes a defense against reactive oxygen species (ROS). TXNRD2 helps facilitate DNA synthesis and repair protein folding and the regulation of apoptosis. As a member of this protective complex TXNRD2 contributes to safeguarding cellular function under various stress conditions by modulating the reduction capacity of cells.

Pathways

TXNRD2 is importantly involved in the antioxidant defense and mitochondrial function pathways. In the antioxidant defense pathway it works in conjunction with the glutathione system to reduce ROS and repair oxidative damage. Additionally in mitochondrial function TXNRD2 collaborates with peroxiredoxins and other mitochondrial antioxidant systems to ensure the integrity of the electron transport chain and prevent oxidative damage within the mitochondria. TXNRD2 shares functional relations with proteins like thioredoxin 1 and peroxiredoxin 3 in these pathways.

TXNRD2 has significant implications in cardiovascular diseases and cancer. Mutations or dysregulation of TXNRD2 can lead to increased oxidative stress and myocardial dysfunction contributing to heart failure. Similarly altered TXNRD2 expression has been associated with tumor progression due to its role in managing cellular apoptosis and antioxidant defense allowing cancer cells to survive in oxidative environments. TXNRD2's interactions with the protein NADPH oxidase highlight its involvement in these conditions where irregular ROS production exacerbates disease progression.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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