Anti-Osteopontin antibody [EPR21138]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] (AB218237)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Osteopontin with ab218237 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in biliary epithelial cells of mouse liver (PMID : 16221502). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] (AB218237)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Osteopontin with ab218237 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in distal tubules of mouse kidney (PMID : 17938278). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody [EPR21138] (AB218237)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labelling Osteopontin with primary antibody anti-Osteopontin (ab218237) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing increased cytoplasmic staining in RAW 264.7 cells treated with LPS (100 ng/ml) for 6 h, then add BFA (300 ng/ml) for 3 h. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue).

• WB

Supplier Data

Western blot - Anti-Osteopontin antibody [EPR21138] (AB218237)

Blocking buffer and concentration : 5% NFDM/TBST. Diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a loading control. This antibody is not recommended for use in WB with mouse tissue samples. We suggest treating cells with BFA which inhibits protein secretion.

All lanes:

Western blot - Anti-Osteopontin antibody [EPR21138] (ab218237) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 4:

RAW 264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 4h whole cell lysate at 20 µg

Lane 5:

RAW 264.7 treated with 1000 ng/ml BFA for 3h whole cell lysate at 20 µg

Lane 6:

RAW 264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 4h then add 1000 ng/ml BFA for another 3h whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 35 kDa

Observed band size: 60 kDa

false

Exposure time: 180s