Human PODXL knockout HeLa cell line
- Advanced Validation
- What is this?
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PODXL KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
GCTM-2 antigen, Gp200, MGC138240, PC, PCLP, PCLP-1, PODXL_HUMAN, Pcx, Podocalyxin, Podocalyxin like, Podocalyxin like protein, Podocalyxin-like protein 1
- WB
Lab
Western blot - Human PODXL knockout HeLa cell line (AB264984)
Lanes 1- 2 : Merged signal (red and green). Green - ab150358 observed at 160 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab150358 was shown to react with PODXL in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264984 (knockout cell lysate ab257210) was used. Wild-type HeLa and PODXL knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab150358 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 10000 dilution and a 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-PODXL antibody [EPR9518] (<a href='/en-us/products/primary-antibodies/podxl-antibody-epr9518-ab150358'>ab150358</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PODXL knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PODXL knockout HeLa cell line (ab264984)
Predicted band size: 58 kDa
Observed band size: 160 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human PODXL knockout HeLa cell line (AB264984)
Allele-2 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PODXL knockout HeLa cell line (AB264984)
Allele-3 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PODXL knockout HeLa cell line (AB264984)
Allele-1 : 1 bp insertion in exon 2.
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, 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
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
The primary function of podocalyxin-like protein revolves around maintaining the filtration barrier in the kidney by repelling negatively charged molecules. It forms part of the slit diaphragm complex along with proteins like nephrin which provides the structural support necessary for its function. PODXL also plays roles in the development of the vascular system and hematopoietic cell lineage contributing to cellular processes like migration and proliferation. Its interaction with the actin cytoskeleton ensures proper localization and function within these biological contexts.
Pathways
There are important connections between podocalyxin-like protein and the Ras signaling pathway involved in cell growth and adhesion processes. It can also engage in integrin-mediated cell adhesion pathways facilitating processes such as cell signaling and migration. Through these pathways it associates with proteins like ezrin which links it to the cytoskeleton and contributes to its role in cell morphology and motility.
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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PODXL antibody [EPR9518] (AB150358)](https://content.abcam.com/products/images/podxl-antibody-epr9518-ab150358--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img64438.jpg)
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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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