rat anti-mouse cd-31  (Santa Cruz Biotechnology)

Journal: Drug Delivery

Article Title: A journey from the endothelium to the tumor tissue: distinct behavior between PEO-PCL micelles and polymersomes nanocarriers

doi: 10.1080/10717544.2018.1510064

Figure Lengend Snippet: Macroscopic and microscopic (cryosection) fluorescent pictures of explanted HCT-116 tumors 24 h after retro-orbital injection in mice of 100 µL of PBS (A,E), DiO alone (B,F), micelles charged with 20% mol DiO (C,G), or polymersomes charged with 20% mol DiO (D,H). DiO fluorescence (green), and in cryosection nuclei stained with Hoechst (blue), and CD-31+ murine endothelial cells detected by immunofluorescence (red).

Article Snippet: Briefly, the slides were rinsed with demineralized water to remove OCT, saturated with MaxBlock Blocking (Active Motif, La Hulpe, Belgium), rinsed and then exposed to anti-CD-31 purified Rat Anti-Mouse primary antibody (BD, Franklin Lakes, USA) at 1/400 in Dako antibody diluent (Dako, Glostrup, Denmark), at 4 °C, overnight.

Techniques: Injection, Fluorescence, Staining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Deletion of Col15a1 Modulates the Tumour Extracellular Matrix and Leads to Increased Tumour Growth in the MMTV-PyMT Mouse Mammary Carcinoma Model

doi: 10.3390/ijms22189978

Figure Lengend Snippet: Mammary carcinogenesis in the wild-type MMTV-PyMT ( PyMT ) and ColXV-deficient PyMT;Col15a1 −/− mice. The incidence ( A ) and number ( B ) of desmoplastic mammary glands (MG) per mouse at the age of 4–13 weeks. Abdominal and thoracic MGs were palpated weekly. ( C ) Total tumour burden in the MGs of PyMT and PyMT;Col15a1 −/− mice at indicated ages, measured as total weight of the dissected four abdominal and two thoracic MGs. The numbers of mice included in the analyses are shown in the bars. ( D ) Representative images of Carmine Alum-stained whole mount preparations of MGs at week 11, and ( E ) haematoxylin-eosin (H&E)-stained MG sections at week 6. ( F ) Representative images of ColXV and CD-31 double immunofluorescence staining in mammary tumours of control PyMT mice at the age of 9 and 12 weeks. ColXV localises in the BMs of tumour nests (arrowheads and magnification in insert) and in vascular BMs, where it often overlaps with the CD-31 signal (arrows). a, adipocyte; s, tumour stroma; t, tumour tissue. Fluorescent signals: ColXV, green; CD-31, red; nuclei, DAPI, blue. The scale bar in E is 200 µm and in F 100 µm. The error bars in B and C indicate s.e.m. * p < 0.05; ** p < 0.01; ns, not significant.

Article Snippet: The following primary antibodies and secondary antibody combinations were used for immunofluorescence staining: polyclonal in-house rabbit ColXV Ab targeting the N-terminal non-collagenous portion of rabbit anti-mouse ColXV (83AA) [ ] at 1:70 dilution, rabbit monoclonal Ki67 Ab (Cat. No. ab15590; Abcam, Cambridge, UK) at a concentration of 0.5 mg/mL with goat anti-rabbit Alexa Fluor 488 (Cat. No. A11008; Invitrogen, Waltham, MA, USA) Ab at 1:200 dilution, monoclonal rat anti-mouse CD-31 Ab (Cat. No. 550274; BD Biosciences, Franklin Lakes, NJ, USA) at 1:100 dilution with goat anti-rat Alexa Fluor 647 Ab (Jackson ImmunoResearch, West Grove, PA, USA) at 1:200 dilution, monoclonal mouse anti-rat PCNA (PC19, sc-56; Santa Cruz, Dallas, Tx, USA) at 1:100 dilution and Cy3-conjugated alpha smooth muscle actin (αSMA) Ab (Cat. No. C6198; Sigma Aldrich) at 1:200 dilution.

Techniques: Staining, Double Immunofluorescence Staining

Journal: International Journal of Molecular Sciences

Article Title: Deletion of Col15a1 Modulates the Tumour Extracellular Matrix and Leads to Increased Tumour Growth in the MMTV-PyMT Mouse Mammary Carcinoma Model

doi: 10.3390/ijms22189978

Figure Lengend Snippet: Angiogenesis and fibrosis in MMTV-PyMT mammary tumours. ( A ) Immunohistochemistry for CD-31 (red) in tumours of 12-week-old control PyMT and PyMT;Col15a1 −/− mice. ( B ) Manual counting of CD-31-positive structures in tumours. For both genotypes ( n = 9), four random microscopic fields were analysed at 100× magnification. ( C ) ImageJ analysis of CD-31-positive areas in the same microscopic fields. Quantification of the signal in the light microscopy images of tumour samples stained with Masson’s trichrome ( D ) and picrosirius red ( E ) revealed comparable fibrillar collagen content in the control and PyMT;Col15a1 −/− tumours. ( F , G ) Quantification of the birefringence of picrosirius red signals captured under polarised light showed a slightly reduced ratio of thick (red) to thin (green) collagen fibres in the PyMT;Col15a1 −/− tumours in comparison to the PyMT control tumours. Samples were collected from mice at 10–14 weeks of age ( n = 8 mice for both genotypes; one tumour sample per mouse was analysed). Four microscopic fields per tumour at 100× magnification were analysed. Scale bars in ( A , D – F ), 200 µm. Error bars in ( B , C , G ) indicate s.e.m. *** p < 0.001; ns, not significant.

Article Snippet: The following primary antibodies and secondary antibody combinations were used for immunofluorescence staining: polyclonal in-house rabbit ColXV Ab targeting the N-terminal non-collagenous portion of rabbit anti-mouse ColXV (83AA) [ ] at 1:70 dilution, rabbit monoclonal Ki67 Ab (Cat. No. ab15590; Abcam, Cambridge, UK) at a concentration of 0.5 mg/mL with goat anti-rabbit Alexa Fluor 488 (Cat. No. A11008; Invitrogen, Waltham, MA, USA) Ab at 1:200 dilution, monoclonal rat anti-mouse CD-31 Ab (Cat. No. 550274; BD Biosciences, Franklin Lakes, NJ, USA) at 1:100 dilution with goat anti-rat Alexa Fluor 647 Ab (Jackson ImmunoResearch, West Grove, PA, USA) at 1:200 dilution, monoclonal mouse anti-rat PCNA (PC19, sc-56; Santa Cruz, Dallas, Tx, USA) at 1:100 dilution and Cy3-conjugated alpha smooth muscle actin (αSMA) Ab (Cat. No. C6198; Sigma Aldrich) at 1:200 dilution.

Techniques: Immunohistochemistry, Light Microscopy, Staining

Journal: International Journal of Molecular Sciences

Article Title: Deletion of Col15a1 Modulates the Tumour Extracellular Matrix and Leads to Increased Tumour Growth in the MMTV-PyMT Mouse Mammary Carcinoma Model

doi: 10.3390/ijms22189978

Figure Lengend Snippet: MCA-induced fibrosarcoma model and syngeneic transplantation models. ( A ) Incidence of MCA-induced subcutaneous fibrosarcomas in Col15a1 +/+ control and Col15a1 −/− knockout mice, presented as the percentage of mice bearing at least one persistent tumour of 3 mm in diameter. ( B ) Tumour volumes at different time points. Numbers of tumours included in the analysis at different time points are shown above the bars. ( C ) Microvessel densities in MCA-induced fibrosarcomas were determined in 10 random fields of CD-31-stained fibrosarcomas ( Col15a1 +/+ n = 12, Col15a1 −/− n = 13) collected 13–17 weeks after MCA treatment at 200× magnification. ( D – F ) Growth curves of subcutaneous syngeneic tumour transplants in the Col15a1 +/+ control and Col15a1 −/− mice. Numbers of mice (N) and tumours (n) included in the experiments are indicated in panels. In ( B – F ), error bars indicate s.e.m.

Article Snippet: The following primary antibodies and secondary antibody combinations were used for immunofluorescence staining: polyclonal in-house rabbit ColXV Ab targeting the N-terminal non-collagenous portion of rabbit anti-mouse ColXV (83AA) [ ] at 1:70 dilution, rabbit monoclonal Ki67 Ab (Cat. No. ab15590; Abcam, Cambridge, UK) at a concentration of 0.5 mg/mL with goat anti-rabbit Alexa Fluor 488 (Cat. No. A11008; Invitrogen, Waltham, MA, USA) Ab at 1:200 dilution, monoclonal rat anti-mouse CD-31 Ab (Cat. No. 550274; BD Biosciences, Franklin Lakes, NJ, USA) at 1:100 dilution with goat anti-rat Alexa Fluor 647 Ab (Jackson ImmunoResearch, West Grove, PA, USA) at 1:200 dilution, monoclonal mouse anti-rat PCNA (PC19, sc-56; Santa Cruz, Dallas, Tx, USA) at 1:100 dilution and Cy3-conjugated alpha smooth muscle actin (αSMA) Ab (Cat. No. C6198; Sigma Aldrich) at 1:200 dilution.

Techniques: Transplantation Assay, Knock-Out, Staining