PODXL KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- 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 lysate available now. KO validated by Western blot. Free of charge wild type control included. 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
- Species or organism
- Human
- Tissue
- Cervix
- 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.
- Disease
- Adenocarcinoma
- Concentration
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
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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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4 product images
Western blot - Human PODXL knockout HeLa cell lysate (ab257210)
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: PODXL knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-PODXL antibody [EPR9518] ab150358 observed at 160 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-PODXL antibody [EPR9518] ab150358 Anti-PODXL antibody [EPR9518] was shown to specifically react with PODXL in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human PODXL knockout HeLa cell line ab264984 (knockout cell lysate ab257210) was used. Wild-type and PODXL knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-PODXL antibody [EPR9518] ab150358 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-PODXL antibody [EPR9518] ab150358) 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 (Human PODXL knockout HeLa cell line ab264984)
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 160 kDa
Sanger Sequencing - Human PODXL knockout HeLa cell lysate (ab257210)
Allele-1: 1 bp insertion in exon 2
Sanger Sequencing - Human PODXL knockout HeLa cell lysate (ab257210)
Allele-2: Insertion of the selection cassette in exon 2
Sanger Sequencing - Human PODXL knockout HeLa cell lysate (ab257210)
Allele-3: Insertion of the selection cassette in exon 2
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