Human P4HB knockout A-431 cell line
- Advanced Validation
- What is this?
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P4HB KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 20 bp deletion Frameshift = 99.4%. 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
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
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-P4HB antibody [EPR9499] (<a href='/en-us/products/primary-antibodies/p4hb-antibody-epr9499-ab137110'>ab137110</a>) 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
Predicted band size: 57 kDa
Observed band size: 60 kDa
false
- NGS
Lab
Next Generation Sequencing - Human P4HB knockout A-431 cell line (AB261887)
X = 20 bp deletion
- NGS
Supplier Data
Next Generation Sequencing - Human P4HB knockout A-431 cell line (AB261887)
20 bp deletion after Leu69 of the WT protein
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Flow Cytometry (Intracellular) - Anti-P4HB antibody [EPR9499] (AB137110)](https://content.abcam.com/products/images/p4hb-antibody-epr9499-ab137110--flow-cytometry-intracellular-img26544.jpg)
Cell Lines & Lysates
AB265841
Human TFG (TRK fused gene) knockout HeLa cell line
Advanced Validation
- 0
- 0
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Primary Antibodies
AB125066
Anti-Glutathione Peroxidase 4 antibody [EPNCIR144]
BOND RX™ Validated|KO Validated|RabMAb|Recombinant|Lab Essentials
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (AB125066)](https://content.abcam.com/products/images/glutathione-peroxidase-4-antibody-epncir144-ab125066--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img436422.jpg)
- 5
- 13
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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