Human PRDX4 (Peroxiredoxin 4) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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PRDX4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
View Alternative Names
AOE37-2, Antioxidant enzyme 372, Antioxidant enzyme AOE372, EC 1.11.1.15, PRDX4_HUMAN, PRX 4, Peroxiredoxin IV, Peroxiredoxin-4, Prx-IV, TRANK, Thioredoxin Peroxidase (Antioxidant Enzyme), Thioredoxin peroxidase, Thioredoxin peroxidase AO372, Thioredoxin-dependent peroxide reductase A0372
- WB
Lab
Western blot - Human PRDX4 (Peroxiredoxin 4) knockout HEK-293T cell line (AB266727)
Lanes 1-3 : Merged signal (red and green). Green - ab184167 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.
ab184167 Anti-Peroxiredoxin 4 antibody [EPR15458(B)] was shown to specifically react with Peroxiredoxin 4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266727 (knockout cell lysate ab257264) was used. Wild-type and Peroxiredoxin 4 knockout samples were subjected to SDS-PAGE. ab184167 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-Peroxiredoxin 4 antibody [EPR15458(B)] (<a href='/en-us/products/primary-antibodies/peroxiredoxin-4-antibody-epr15458b-ab184167'>ab184167</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
PRDX4 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human PRDX4 (Peroxiredoxin 4) knockout HEK-293T cell line (ab266727)
Lane 3:
HepG2 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: 177 kDa,189 kDa,31 kDa
Observed band size: 27 kDa
false
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
Prdx4 plays a list of roles in cellular defense against oxidative stress. It not only reduces peroxides but also contributes to the maintenance of protein folding by modifying disulfide bonds in the endoplasmic reticulum. Prdx4 does not function as part of a larger complex but acts independently within its compartments. The protective functions of Prdx4 are important for cell survival and proper cellular function.
Pathways
Prdx4 interacts with oxidative stress response pathways. Specifically it participates in the redox signaling pathways that regulate cellular reactive oxygen species levels. Peroxiredoxin 4 is closely associated with proteins such as thioredoxin in these pathways. Together they regulate the cellular redox environment which is important for maintaining cellular homeostasis.
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
Complementary Products
Primary Antibodies
AB184167
Anti-Peroxiredoxin 4 antibody [EPR15458(B)]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-Peroxiredoxin 4 antibody [EPR15458(B)] (AB184167)](https://content.abcam.com/products/images/peroxiredoxin-4-antibody-epr15458b-ab184167--western-blot-img105043.png)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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