Anti-Podoplanin抗体[18H5] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

Immunocytochemistry/Immunofluorescence analysis of human dermal lymphatic endothelial cells (HDLEC) labelling Podoplanin with ab10288. An Alexa Fluor^® 488-conjugated goat anti-mouse was used as the secondary antibody. Nuclei counterstained with DAPI.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

Flow cytometric analysis of primary human dermal microvascular (HDMVEC) and umbilical vein (HUVEC) endothelial cells labeling Podoplanin with ab10288 at 5 μg/mL. The lymphatic endothelial cell marker Podoplanin is not expressed in the blood endothelial cells.

• Flow Cyt

Unknown

Flow Cytometry - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

Overlay histogram showing HCT116 cells stained with ab10288 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10288, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HCT116 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

• ICC/IF

AbReview4632****

Immunocytochemistry/ Immunofluorescence - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

ab10288 at a 1/500 dilution staining human dermal lymphatic endothelial by Immunocytochemistry. The cells were grown in coverslips and fixed in formalin prior to permeabilization with tritonX-100. The cells were incubated with the primary antibody at 1/500 for 1 hour and then bound antibody was detected using a biotinylated goat anti-mouse antibody. Texas red-streptavidin was used for development.

This image is courtesy of an Abreview submitted by Maria Navarro.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

Flow cytometric analysis of human primary lymphatic endothelial cells (HDLEC) labeling Podoplanin with ab10288 at 5 μg/mL. Anti-mouse IgG-PE was used as a secondary antibody.

• WB

Unknown

Western blot - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

Podoplanin protein is extensively glycosylated. This might explain the observation of a band at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-Podoplanin antibody [18H5] - BSA and Azide free (ab10288) at 5 µg/mL

Lane 1:

Human placenta tissue lysate - total protein (<a href='/products/unavailable/human-placenta-tissue-lysate-total-protein-ab29745'>ab29745</a>) at 20 µg

Lane 2:

Human skeletal muscle tissue lysate - total protein (<a href='/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 16 kDa

Observed band size: 35 kDa

true

Exposure time: 20min

• WB

Supplier Data

Western blot - Anti-Podoplanin antibody [18H5] - BSA and Azide free (AB10288)

Western blot analysis of Podoplanin expression in human lymphatic endothelial cells (LEC) - lane 1, and HUVECs - lane 2. Total lysate of both cell types were subjected to SDS-PAGE and subsequent western blot analysis with ab10288. The antibody recognizes a protein of about 36 kDa in total lysate from LECs but not from HUVEC.

All lanes:

Western blot - Anti-Podoplanin antibody [18H5] - BSA and Azide free (ab10288)

Predicted band size: 16 kDa

false