Cancer Research Technology
Log in Register
Menu
Search
Monoclonal Antibodies

Anti-Myofibroblast marker [PR 2D3]

Invented by Prof Walter Bodmer from University of Oxford
Invented at Cancer Research UK London Research Institute: Lincoln's Inn Fields
  • Datasheet
  • Inventor Info
Print

Info

Image thumbnail for Anti-Myofibroblast marker [PR 2D3]
Image thumbnail for Anti-Myofibroblast marker [PR 2D3]
Add an image
Adapted from Hsia LT, Ashley N, Ouaret D, Wang LM, Wilding J, Bodmer WF. Myofibroblasts are distinguished from activated skin fibroblasts by the expression of AOC3 and other associated markers. Proc Natl Acad Sci U S A. 2016 Apr 12;113(15):E2162-71. doi: 10.1073/pnas.1603534113. Epub 2016 Mar 28. PMID: 27036009; PMCID: PMC4839407. AOC3 is expressed in primary MFs but not in fibroblasts. (A) Immunofluorescent staining of CCD 18CO cells and foreskin fibroblast cells with AOC3 mAb (TK8-14) and PR2D3. (Magnification: 20×.) (B) Western blot using AOC3 mAb (TK8-14) detected a 150-kDa protein in CCD 18CO, myo2020, myo1998, and myo6544 cells but not in foreskin fibroblasts (F. Fibro) or skin fibroblasts (Skin Fibro). Tubulin was used as the loading control. (C) Flow cytometric analysis of primary cell cultures (myo6544 and myo1998) and CCD 18CO cells. Fibroblasts (foreskin fibroblasts and two sets of adult skin fibroblasts) and epithelial cells (SW1222 and LS174T) show that MFs are AOC3+, whereas fibroblasts and epithelial cells do not express AOC3. Isotype control is shown in red, and anti-AOC3 (TK8-14) is shown in green. (D) mRNA-expression levels of AOC3 and ACTA2 in MF and fibroblast cultures. AOC3 and ACTA2 mRNA expression in eight cell lines (four MF cultures and four fibroblast cultures) was measured by the Affymetrix Human Genome U133 Plus 2.0 microarray. The mRNA-expression levels along the y axis are actual ΔCt fluorescence intensities. (E) AOC3 functions as an SSAO enzyme in MFs. The enzyme activity of MFs was determined by SSAO-mediated H2O2 production. The siRNA for AOC3, the SSAO inhibitor semicarbazide (SEM) (1 mM), and the MAO-A inhibitor Clorgyline (1 mM) were added under serum-free conditions for 48 h. The enzyme activity in untreated CCD 18CO control cells was set to 100%. The P values are based on t tests for the difference between the treated and the serum-free control samples: **P < 0.001.
Adapted from Hsia LT, Ashley N, Ouaret D, Wang LM, Wilding J, Bodmer WF. Myofibroblasts are distinguished from activated skin fibroblasts by the expression of AOC3 and other associated markers. Proc Natl Acad Sci U S A. 2016 Apr 12;113(15):E2162-71. doi: 10.1073/pnas.1603534113. Epub 2016 Mar 28. PMID: 27036009; PMCID: PMC4839407. The MF marker mAb PR2D3 recognizes the AOC3 protein in human colonic lysates. (A) Immunofluorescence staining of cryostat sections of normal colon and CRC tissue using PR2D3 (green) for MFs and AUA-1 (red) to identify epithelial cells. (B) The immunoblot of human colonic smooth muscle (SM) lysate using mAb PR2D3 shows a band of about 150 kDa in a native (nonreduced) sample.
Add an image
Image thumbnail for Anti-Myofibroblast marker [PR 2D3] Image thumbnail for Anti-Myofibroblast marker [PR 2D3]
Catalogue Number 151152
Applications IHC IF IP WB
Antigen/Gene or Protein Targets Myofibroblast marker
Reactivity Human
Relevance PR 2D3 reacts with a cell membrane component of cells in the pericrypt sheath; with smooth muscle cells and myofibroblasts. This antibody is considered to be the gold standard for the identification of myofibroblasts.
Host Mouse
Immunogen Crude homogenate of normal colorectal mucosa.
Subclass IgG1
Molecular Weight (kDa) 140
Myeloma Used NS0
Recommended Growing Conditions RPMI 1640 + 10% FCS.
Strain Balb/c
Research Area Cell Type or Organelle Marker

References

There are 6 reference entries for this reagent.

View All References

References: 6 entries

Hsia et al. 2016. Proc Natl Acad Sci U S A. :. PMID: 27036009.

IP WB

View  

Myofibroblasts are distinguished from activated skin fibroblasts by the expression of AOC3 and other associated markers.

Europe PMC ID: 27036009

View  

Kirkland et al. 2001. J Cell Sci. 114(Pt 11):2055-64. PMID: 11493641.

View  

Collagen IV synthesis is restricted to the enteroendocrine pathway during multilineage differentiation of human colorectal epithelial stem cells.

Europe PMC ID: 11493641

View  

Richman et al. 1987. J Clin Pathol. 40(6):593-600. PMID: 3301906.

View  

Colonic pericrypt sheath cells: characterisation of cell type with new monoclonal antibody.

Europe PMC ID: 3301906

View  

Add a reference

References: 6 entries

Hsia et al. 2016. Proc Natl Acad Sci U S A. :. PMID: 27036009.

IP WB

View  

Myofibroblasts are distinguished from activated skin fibroblasts by the expression of AOC3 and other associated markers.

View  

Kirkland et al. 2001. J Cell Sci. 114(Pt 11):2055-64. PMID: 11493641.

View  

Collagen IV synthesis is restricted to the enteroendocrine pathway during multilineage differentiation of human colorectal epithelial stem cells.

View  

Richman et al. 1987. J Clin Pathol. 40(6):593-600. PMID: 3301906.

View  

Colonic pericrypt sheath cells: characterisation of cell type with new monoclonal antibody.

View  

Add a reference

Inventor Information