P4HB KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 20 bp deletion Frameshift = 99.4%.
Images
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 20 bp deletion Frameshift = 99.4%
Alternative names
Cellular thyroid hormone-binding protein, Collagen prolyl 4 hydroxylase beta, DSI, Disulphide Isomerase, EC 5.3.4.1, ER protein 59, ERBA2L, ERp59, Endoplasmic reticulum resident protein 59, GIT, Gltathione insulin transhydrogenase, Glutathione insulin transhydrogenase, P4Hbeta, PDIA1_HUMAN, PDIR, PHDB, PO4DB, PO4HB, PROHB, Procollagen proline 2 oxoglutarate 4 dioxygenase (proline 4 hydroxylase) beta polypeptide (protein disulfide isomerase associated 1), Procollagen proline 2 oxoglutarate 4 dioxygenase beta subunit, Prolyl 4 hydroxylase beta polypeptide, Prolyl 4 hydroxylase beta subunit, Prolyl 4-hydroxylase subunit beta, Protein disulfide isomerase associated 1, Protein disulfide isomerase, family A, member 1, Protein disulfide isomerase/oxidoreductase, Protein disulfide-isomerase, Protocollagen hydroxylase, Thbp, Thyroid hormone binding protein p55, Thyroid hormone binding protein p55 cellular, V erb a avian erythroblastic leukemia viral oncogene homolog 2 like, p55
Recommended products
P4HB KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 20 bp deletion Frameshift = 99.4%.
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 20 bp deletion Frameshift = 99.4%
- Disease
- Epidermoid Carcinoma
- Concentration
Properties
- Gene name
- P4HB
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
P4HB is a protein known as Protein Disulfide Isomerase (PDI). It plays a significant role in catalyzing the formation of disulfide bonds. This protein helps to fold newly synthesized proteins by shuffling disulfide bonds. It has a molecular mass of around 57 kDa. P4HB is mainly expressed in the endoplasmic reticulum of cells. While its alternative name is ERp59 P4HB is integral to the protein folding machinery inside cells.
- Biological function summary
P4HB functions in protein folding and assembly. By acting as a chaperone it protects proteins from misfolding and aggregation. P4HB operates as part of a larger multi-protein complex that assists in maintaining protein structure under stress conditions in the cell. Its activity ensures protein stability and proper cellular function important for cell viability and health.
- Pathways
P4HB plays a central role in the unfolded protein response (UPR) and oxidative protein folding pathway. It interacts with proteins such as calnexin and calreticulin through its involvement in these pathways. During oxidative protein folding P4HB introduces disulfide bonds into nascent proteins while removing incorrect ones ensuring efficient protein quality control.
- Associated diseases and disorders
P4HB alteration has connections to diseases like cancer and neurodegenerative disorders. Its overexpression and activity can promote tumor growth by aiding cancerous cell survival through protein homeostasis. Moreover P4HB's association with amyloid precursor protein impacts the progression of Alzheimer's disease. These connections make P4HB a relevant target for research into therapeutic interventions.
Product promise
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Western blot - Human P4HB knockout A-431 cell line (ab261887)
False colour image of Western blot: Anti-P4HB antibody [EPR9499] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-P4HB antibody [EPR9499] ab137110 was shown to bind specifically to P4HB. A band was observed at 60 kDa in wild-type HeLa cell lysates with no signal observed at this size in P4HB knockout cell line ab261887 (knockout cell lysate Human P4HB knockout A-431 cell lysate ab261696). To generate this image wild-type and P4HB knockout HeLa cell lysates were analysed. First 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°. 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-P4HB antibody [EPR9499] (Anti-P4HB antibody [EPR9499] ab137110) at 1/1000 dilution
Lane 1: HeLa cell lysate at 20 µg
Lane 2: HepG2 cell lysate at 20 µg
Lane 2: Western blot - Human P4HB knockout A-431 cell line (ab261887)
Lane 3: Wild-type A431 cell lysate at 20 µg
Lane 4: P4HB knockout A431 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 60 kDa
Next Generation Sequencing - Human P4HB knockout A-431 cell line (ab261887)
X = 20 bp deletion
Next Generation Sequencing - Human P4HB knockout A-431 cell line (ab261887)
20 bp deletion after Leu69 of the WT protein
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