This is a knockout-validated antibody summary, based on the publications "Hic-5 remodeling of the stromal matrix promotes breast tumor progression", as cited below [1] and "The focal adhesion scaffold protein Hic-5 regulates vimentin organization in fibroblasts" for immunocytochemistry knockout validation (see figure 3a [2] ). Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG1

Company: BD Biosciences

Antibody: Hic5

Catalog number: 611164

Summary: Mouse monoclonal IgG1 against mouse Hic-5 (aa. 73-186). Reacts with mouse, rat, human and dog. Suitable for western blot and immunofluorescence.

Western blot | Immunohistochemistry | Immunocytochemistry

Tumors and cancer-associated fibroblasts from Hic-5+/− ;PyMT or Hic-5−/− ;PyMT mice. IHC: Tumor sections from endpoint mice were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS), embedded in OCT and sectioned. IC: Cells were washed once in PBS, fixed, and permeabilized simultaneously using 4% paraformaldehyde with 1% Triton X-100 in PBS and quenched with 0.1 M glycine in PBS.

IHC: 10% normal goat serum for 2 h.

IC: 3% (wt/vol) BSA overnight at 4°C.

IHC: diluted in 10% normal goat serum.

IHC: for 2 h.

IHC: Zeiss Axioskop2 plus microscope fitted with a Q imaging EXi Blue CCD camera using a Plan-Apochromat 20 × /0.75 NA objective.

IC: Leica SP5 scanning confocal with a HCX PL APO 63×/1.40–0.60 OIL λ BL objective (Leica, Bannockburn, IL).

Representative western blots of Hic-5 in tumors (c) and cancer-associated fibroblasts (i) from Hic-5+/− ;PyMT or Hic-5−/− ;PyMT mice. (e) Representative immunohistochemistry of Hic-5 staining in tumors from endpoint PyMT mice. The inset shows Hic-5 staining in the stroma (S), but not in the tumor cells (T). n =3 mice of each genotype. (g) Representative staining of Hic-5 and paxillin in isolated CAFs. n =3 independent experiments. Please see Figure 1 in the article [1].