Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell line
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PLOD2 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.
View Alternative Names
2-oxoglutarate 5-dioxygenase 2, LH2, Lysine hydroxylase 2, Lysyl hydroxylase 2, OTTHUMP00000215204, OTTHUMP00000215205, OTTHUMP00000215206, PLOD2_HUMAN, Procollagen lysine 2 oxoglutarate 5 dioxygenase 2, Procollagen lysine, 2 oxoglutarate 5 dioxygenase (lysine hydroxylase) 2, Procollagen-lysine, TLH, Telopeptide lysyl hydroxylase
- WB
Supplier Data
Western blot - Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell line (AB266279)
Blocking and diluting buffer and concentration : 5% NFDM/TBST
Lysates at 20 µg per lane.
The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-PLOD2/LH2 antibody (ab313765) 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, ab313765 was shown to bind specifically to PLOD2/LH2.
Target of interest was observed at 95 kDa in wild-type Hela cell lysates (lane 1) with no signal observed at this size in PLOD2 knockout cell line ab266279 (knockout cell lysate ab258591) (lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
All lanes:
Western blot - Anti-PLOD2/LH2 antibody [EPR25160-20] (<a href='/en-us/products/primary-antibodies/plod2-lh2-antibody-epr25160-20-ab313765'>ab313765</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell line (ab266279)
Lane 2:
Western blot - Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-plod2-plod2-lh2-knockout-hek-293t-cell-lysate-ab258591'>ab258591</a>) at 20 µg
Lane 3:
HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4:
A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution
Observed band size: 95 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PLOD2 (PLOD2/LH2) knockout HEK-293T cell line (AB266279)
Homozygous : 1 bp insertion in exon4
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.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com