Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

Immunohistochemical analysis of paraffin-embedded human chondrosarcoma tissue labeling Sp7 / Osterix with ab209484 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on tumor cells of human chondrosarcoma is observed. 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 (ab209484).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

This data was developed using ab209484, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human chondrosarcoma labelling Sp7/Osterix with ab209484 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab209484 anti-Sp7/Osterix antibody [EPR21034] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IP

Supplier Data

Immunoprecipitation - Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

Sp7 / Osterix was immunoprecipitated from 0.35mg of Saos-2 (human osteosarcoma cell line) whole cell lysate with ab209484 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209484 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Saos-2 whole cell lysate 10ug (Input).

Lane 2 : ab209484 IP in Saos-2 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab190908 in Saos-2 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209484).

All lanes:

Immunoprecipitation - Anti-Sp7 / Osterix antibody [EPR21034] (<a href='/en-us/products/primary-antibodies/sp7-osterix-antibody-epr21034-ab209484'>ab209484</a>)

Predicted band size: 45 kDa,47 kDa

Observed band size: 45 kDa,47 kDa

false

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse E14.5 developing humerus tissue labeling Sp7 / Osterix with ab209484 at 1/500 dilution, followed by AlexaFluor^® 488 Goat anti-Rabbit (ab150077) at 1/1000 dilution. Positive staining on the mouse embryo E14.5 developing humerus (PMID : 27134141) is observed. Counter stained with DAPI.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor^® 488 Goat anti-Rabbit (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209484).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

Immunohistochemical analysis of paraffin-embedded rat E14.5 rib tissue labeling Sp7 / Osterix with ab209484 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on osteoblasts and chondrocytes of rat E14.5 rib (PMID : 17579353; PMID : 25977369) is observed. 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 (ab209484).

Perform heat mediated antigen retrieval 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-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

Immunohistochemical analysis of paraffin-embedded mouse E14.5 rib tissue labeling Sp7 / Osterix with ab209484 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on osteoblasts and chondrocytes of mouse E14.5 rib (PMID : 17579353; PMID : 25977369) is observed. 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 (ab209484).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Sp7 / Osterix antibody [EPR21034] - BSA and Azide free (AB227820)

Blocking and dilution buffer concentration : 5% NFDM/TBST.

We recommend using 1% SDS hot method to prepare lysate to improve signal intensity. Please refer to the Protocol booklet under the Support & downloads section.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209484).

All lanes:

Western blot - Anti-Sp7 / Osterix antibody [EPR21034] (<a href='/en-us/products/primary-antibodies/sp7-osterix-antibody-epr21034-ab209484'>ab209484</a>) at 1/1000 dilution

Lane 1:

Saos-2 (human osteosarcoma cell line) whole cell lysate, prepared by RIPA method at 20 µg

Lane 2:

Saos-2 (human osteosarcoma cell line) whole cell lysate, prepared by 1% SDS hot method at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa,47 kDa

false

Exposure time: 20s