Overview

  • Product name

    Anti-Sp7 / Osterix antibody [EPR21034]
    See all Sp7 / Osterix primary antibodies
  • Description

    Rabbit monoclonal [EPR21034] to Sp7 / Osterix
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Fr, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse Sp7/ Osterix aa 1-250. The exact sequence is proprietary.
    Database link: Q8VI67

  • Positive control

    • WB: Saos-2 whole cell lysate. IHC: Mouse E14.5 rib, rat E14.5 rib and human chondrosarcoma tissue. IHC-Fr: Mouse embryo E14.5 developing humerus tissue. IP: Saos-2 whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209484 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 45, 47 kDa (predicted molecular weight: 45 kDa).
IP 1/30.
IHC-Fr 1/500.

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

IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

Images

  • Anti-Sp7 / Osterix antibody [EPR21034] (ab209484) at 1/1000 dilution + Saos-2 (human osteosarcoma cell line) whole cell lysate at 10 µg

    Secondary
    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, 47 kDa
    why is the actual band size different from the predicted?



    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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

References

This product has been referenced in:

  • Studentsova V  et al. Insulin Receptor deletion in S100a4-lineage cells accelerates age-related bone loss. Bone Rep 10:100197 (2019). Read more (PubMed: 30805422) »
  • Wang C  et al. Poly(ADP-ribose) polymerase 1 accelerates vascular calcification by upregulating Runx2. Nat Commun 10:1203 (2019). Read more (PubMed: 30867423) »
See all 6 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Osteoblasts)
Antigen retrieval step
None
Permeabilization
Yes - 20%DMSO
Specification
Osteoblasts
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 30 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Embryonic bone)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Embryonic bone
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Feb 11 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Embryo)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Embryo
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Feb 11 2019

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