Anti-Sp7 / Osterix antibody [EPR21034]

9 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  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.

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  IHC image of SP7/Osterix staining in a section of formalin-fixed paraffin-embedded normal rat E15 embryo performed on a Leica BOND® system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with Anti-Sp7 / Osterix antibody ab22552, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
  

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  IHC image of SP7/Osterix staining in a section of formalin-fixed paraffin-embedded normal mouse E15 embryo performed on a Leica BOND® system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with Anti-Sp7 / Osterix antibody ab22552, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
  

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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  Western blot - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  Human Sp7 / Osterix has two isoforms (45 and 47 kDa). The 45kDa isoform is predominant (UniProt ID: Q8TDD2). In Saos-2 cells, we observed a faint, extra band at around 37kDa.

  All lanes: Western blot - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484) at 1/1000 dilution

  All lanes: Saos-2 (human osteosarcoma cell line) whole cell lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 45 kDa

  Observed band size: 45 kDa, 47 kDa

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  Immunohistochemistry (Frozen sections) - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  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.

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

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  Immunoprecipitation - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-KAT7 / Hbo1 / MYST2 antibody [EPR18473] ab190908 in Saos-2 whole cell lysate.

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

  All lanes: Immunoprecipitation - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  Predicted band size: 45 kDa, 47 kDa

  Observed band size: 45 kDa, 47 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  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.

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

• 

  Western blot - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484)

  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.

  All lanes: Western blot - Anti-Sp7 / Osterix antibody [EPR21034] (ab209484) 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 45 kDa

  Observed band size: 45 kDa, 47 kDa

  Exposure time: 20s