Podocalyxin Like Antibody - BSA Free

Novus Biologicals | Catalog # NB110-41503

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Podocalyxin Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide within an internal region (residues 100-200) of the human Podocalyxin protein. [Swiss-Prot: O00592] This immunogen should be far enough away from any glycosylation site.

Localization

Membrane

Marker

Podocytes /Apical Marker, Endothelial Cell Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Podocalyxin Like Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Podocalyxin Like Antibody [NB110-41503]

Immunocytochemistry/ Immunofluorescence: Podocalyxin Like Antibody [NB110-41503]

Immunocytochemistry/Immunofluorescence: Podocalyxin Like Antibody [NB110-41503] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-Podocalyxin Like Antibody at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Applications for Podocalyxin Like Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:1000

Western Blot

2 ug/ml
Application Notes
In Western blot, a band is observed at ~52 kDa. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. In ICC/IF membrane staining was observed.

Reviewed Applications

Read 1 review rated 3 using NB110-41503 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: Podocalyxin

Podocalyxin (PODXL) is an extensively N/O-glycosylated and sialylated type I transmembrane protein that contains C-terminal PDZ-binding motif DTHL capable of facilitating interactions with NHERF1/NHERF2 adaptor proteins which complexes with proteins implicated in ion transport, protein trafficking and signal transduction. PODXL function as an anti-adhesive molecule that maintains an open filtration pathway between neighboring foot processes in podocytes by charge repulsion. PODXL also acts as a pro-adhesive molecule, enhancing the adherence of cells to immobilized ligands, increasing the rate of migration and cell-cell contacts in an integrin-dependent manner. It induces apical actin-dependent microvilli formation and is implicated preapical plasma membrane subdomain formation to set up inital epithelial polarization and apical lumen formation during renal tubulogenesis. PODXL is highly expressed in renal glomeruli, embryonic germ layers, hematopoietic progenitors, megakaryocytes and platelets, vascular endothelia, mesothelial cells lining organs, and neurons. In several malignancies such as leukemia and cancer of breast, testicular, prostate etc, PODXL interactions with actin-binding protein EZR enhances cancer aggressiveness by inducing cell migration and invasion through via modulation of MAPK/PI3K signaling.

Alternate Names

GCTM, PCLP, POD XL, PODXL

Gene Symbol

PODXL

Additional Podocalyxin Products

Product Documents for Podocalyxin Like Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for Podocalyxin Like Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Podocalyxin Like Antibody - BSA Free

Customer Reviews for Podocalyxin Like Antibody - BSA Free (1)

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  • Podocalyxin Like Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: lysates from melanoma cells
    Species: Human
    Verified Customer | Posted 08/21/2017
    Tested on Peggy Sue. I used lysates from melanoma cells, melanoma extracellular vesicles (EV) and lysates from melanoma exosomes.
    The polyclonal antibody NB110-41503 yields a very intense signal with several peaks. The peaks are all around the expected size, but I don’t know if the multiple peaks are due to the polyclonal nature of the antibody or different proteins being detected. The signal is absent in the exosome lysate, in line with the observation made using antibody MAB1658.
    Podocalyxin Like Antibody - BSA Free NB110-41503

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Protocols

View specific protocols for Podocalyxin Like Antibody - BSA Free (NB110-41503):

Podocalyxin Like Antibody:
Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Podocalyxin Like Antibody:
Western Blot Protocol

1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 30 ug of total protein per
lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS, 1 hour at room temperature.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-Podolyxin primary antibody (NB 110-41503) in blocking buffer and incubate 2 hours at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers
instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce, ECL).

Note: Tween-20 can be added to the blocking or antibody diultion buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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