Anti-Osteopontin 抗体 [EPR21138] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854)

This data was developed using ab218237, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Liver tissue from wild-type C57BL/6J mice; (B) Liver tissue from Spp1 (Osteopontin) knockout mice tissue labeling with ab218237 at 1/2000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on biliary epithelial cells of (A) Liver tissue from wild-type C57BL/6J mice, no staining on (B) Liver tissue from Spp1 (Osteopontin) knockout mice.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

The tissue samples were kindly provided by GemPharmatech. C57BL/6 wildtype mice and Spp1-KO homozygous mice (Strain ID : T064052).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854)

This data was developed using ab218237, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Kidney tissue from wild-type C57BL/6J mice; (B) Kidney tissue from Spp1 (Osteopontin) knockout mice tissue labeling with ab218237 at 1/2000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on biliary epithelial cells of (A) Kidney tissue from wild-type C57BL/6J mice, no staining on (B) Kidney tissue from Spp1 (Osteopontin) knockout mice.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

The tissue samples were kindly provided by GemPharmatech. C57BL/6 wildtype mice and Spp1-KO homozygous mice (Strain ID : T064052).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218237).

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] - BSA and Azide free (AB229854)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218237).

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] - BSA and Azide free (AB229854)

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). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218237).

• WB

Supplier Data

Western blot - Anti-Osteopontin antibody [EPR21138] - BSA and Azide free (AB229854)

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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218237).

All lanes:

Western blot - Anti-Osteopontin antibody [EPR21138] (<a href='/products/primary-antibodies/osteopontin-antibody-epr21138-ab218237'>ab218237</a>) 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='/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