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PODXL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.

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Images

Western blot - Human PODXL knockout HeLa cell line (AB264984), expandable thumbnail
  • Sanger Sequencing - Human PODXL knockout HeLa cell line (AB264984), expandable thumbnail
  • Sanger Sequencing - Human PODXL knockout HeLa cell line (AB264984), expandable thumbnail
  • Sanger Sequencing - Human PODXL knockout HeLa cell line (AB264984), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2

Alternative names

GCTM-2 antigen, Gp200, MGC138240, PC, PCLP, PCLP-1, PODXL_HUMAN, Pcx, Podocalyxin, Podocalyxin like, Podocalyxin like protein, Podocalyxin-like protein 1

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PODXL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
PODXL
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 2 US: 2
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

Recommended control: Human wild-type HeLa cell line (ab255448). 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

PODXL also known as podocalyxin-like protein is a type I transmembrane glycoprotein possessing a mass of approximately 165 kDa. This protein is a member of the CD34 family and exhibits expression primarily on the apical surface of podocytes in the kidney most hematopoietic stem cells and certain cancer cell types. In its mechanical role PODXL mainly functions in cell adhesion and the maintenance of cell shape through its cytoplasmic domain that connects with the cytoskeleton. The sialylated and sulfated glycosaminoglycan chains on the extracellular domain contribute to its charge and barrier functions.

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.

Associated diseases and disorders

Altered PODXL expression shows relevance in aggressive forms of cancer including renal cell carcinoma and breast cancer. The protein associates with different markers of poor prognosis making it of interest in cancer research. Furthermore in kidney diseases such as focal segmental glomerulosclerosis abnormal PODXL expression affects the slit diaphragm's function leading to proteinuria. Its interaction with nephrin can highlight disrupted cellular architecture typical of these disease states.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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