| Application | Recommended Dilution |
|---|---|
| WB | 1:500-1:5000 |
| IHC | 1:50-1:500 |
| IF | 1:50-1:200 |
| IP | 1:200-1:2000 |
Western Blot
Positive WB detected in: Hela whole cell lysate, HEK293 whole cell lysate
All lanes: PODXL antibody at 2.5µg/ml
Secondary
Goat polyclonal to Mouse IgG at 1/5000 dilution
Predicted band size: 59, 56 kDa
Observed band size: 59 kDa
Western Blot
Positive WB detected in: A549 whole cell lysate
All lanes: PODXL antibody at 2.5µg/ml
Secondary
Goat polyclonal to Mouse IgG at 1/5000 dilution
Predicted band size: 59, 56 kDa
Observed band size: 59 kDa
Western Blot
Positive WB detected in: Mouse brain tissue
All lanes: PODXL antibody at 0.43µg/ml
Secondary
Goat polyclonal to Mouse IgG at 1/5000 dilution
Predicted band size: 59, 56 kDa
Observed band size: 59 kDa
Western Blot
Positive WB detected in: Rat brain tissue, Rat heart tissue, Rat kidney tissue
All lanes: PODXL antibody at 1.28µg/ml
Secondary
Goat polyclonal to Mouse IgG at 1/5000 dilution
Predicted band size: 59, 56 kDa
Observed band size: 59 kDa
Immunohistochemistry of paraffin-embedded human kidney tissue using CSB-MA134471A0m at dilution of 1:200
Immunohistochemistry of paraffin-embedded human melanoma using CSB-MA134471A0m at dilution of 1:200
Immunofluorescent analysis of Hela cells using CSB-MA134471A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Immunofluorescent analysis of PC-3 cells using CSB-MA134471A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Immunofluorescent analysis of A549 cells using CSB-MA134471A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Immunofluorescent analysis of Ntera-2 cells using CSB-MA134471A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
Overlay histogram showing Hela cells stained with CSB-MA134471A0m (red line). The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (10µg/1*106cells) for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was mouse IGg2b (10µg/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
Immunoprecipitating PODXL in HEK293 whole cell lysate
Lane 1: Mouse control IgG (1µg) instead of CSB-MA134471A0m in HEK293 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-MA134471A0m (8µg) + HEK293 whole cell lysate (500µg)
Lane 3: HEK293 whole cell lysate (10µg)
The PODXL (podocalyxin-like) antibody is a monoclonal antibody identified as the IgG2b isotype. Its production begins with immunizing mice with the recombinant human PODXL protein, isolating the PODXL-producing B cells, then fusing the PODXL-producing B cells with myeloma cells, injecting the PODXL-producing hybridoma cells into the abdominal cavity of mice, and finally collecting mouse ascites. This PODXL antibody was purified from mouse ascites fluids by protein G. Its purification reaches up to 95%. It can react with samples containing the PODXL protein from humans, mice, and rats. The quality of this PODXL monoclonal antibody has been tested in ELISA, WB, IHC, IP, IF, and FC applications.
PODXL, a highly glycosylated type I transmembrane protein associated with CD34, is highly expressed in glomerular podocytes, vascular endothelium, hematopoietic cells, and breast epithelial cells. The high expression of PODXL in these tissues initiates the occurrence of many physiologic processes, such as hematopoiesis, leucocyte-endothelial cell interaction, as well as the regulation of vascular permeability and neural development. PODXL is also involved in the modulation of adhesion, cell morphology, and cancer progression.
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