Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Characterization of PRLR isoform antibodies. A. Western blot analysis indicating specificity of polyclonal antibodies. CHO cells were transiently transfected with isoform specific PRLR cDNA as described. Cell lysates were prepared and proteins separated by PAGE. Each isoform specific lysate was probed with each isoform specific antibody. Each transfection was performed in triplicate and Western blot analysis performed twice. Data shown are a representative autoradiogram. Molecular weights are marked as indicated: LF = 93 kDa, SF1a = 57 kDa, SF1b = 38 kDa. Cell lysates from CHO transfected cells were immunoprecipitated with their respective antibodies, then immunoblotted with the same antibodies. Each transfection was performed in triplicate and Western blot analysis performed twice. Data shown are a representative autoradiogram. IP indicates immunoprecipitation. B. Western blot analysis using commercially available PRLR antibody. ECD = antibody recognizing extracellular domain C. Fluorescent immunocytochemical analysis. CHO cells were transiently transfected with isoform specific PRLR cDNA as described. Specific staining was observed using isoform specific polyclonal antibodies. The negative control (not shown) lacks primary antibody and appears black. Data shown are representative of triplicate experiments. Magnification 20×.

Article Snippet: After blocking in 5% normal goat serum (Jackson Laboratories, Bar Harbor, ME) prepared in PBS-0.1% Triton, slides were incubated with the PRLR isoform specific antibodies (10 μg/ml) for 2 hr at room temperature.

Techniques: Western Blot, Transfection, Immunoprecipitation, Staining, Negative Control

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Immunoanalysis on human xenografts. A. Fluorescent immunohistochemistry was performed on MCF7, MDA-MB-231, or T47D cells which were injected into the cleared fat pads of nude mice; once tumors reached 1 cm 2 (measured in two dimensions), they were excised, formalin fixed, sectioned, and stained with the indicated PRLR isoform specific antibodies. B. Immunohistochemical analysis using DAB as chromogen of MCF7, MDA-MB-231, or T47D xenograft tumors. neg = no primary antibody as negative control; DAPI stain to visualize the presence of cells. Photographs are representative from 5 different tumors for each cell line. Magnification 40×.

Article Snippet: After blocking in 5% normal goat serum (Jackson Laboratories, Bar Harbor, ME) prepared in PBS-0.1% Triton, slides were incubated with the PRLR isoform specific antibodies (10 μg/ml) for 2 hr at room temperature.

Techniques: Immunohistochemistry, Injection, Staining, Immunohistochemical staining, Negative Control

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Message for PRLR isoforms in breast carcinomas. RNA was extracted from 7 ductal (left) and 7 lobular (right) carcinomas and qPCR was performed. Data shown are the mean +/- SE of delta delta Ct values relative to SF1a expression.

Article Snippet: After blocking in 5% normal goat serum (Jackson Laboratories, Bar Harbor, ME) prepared in PBS-0.1% Triton, slides were incubated with the PRLR isoform specific antibodies (10 μg/ml) for 2 hr at room temperature.

Techniques: Expressing

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Immunohistochemical analysis from ductal breast tissue arrays. Representative photographs taken of ductal carcinoma specimens to demonstrate absent, low, medium, or high amounts of fluorescent red intensity staining. 144 individual ductal specimens were stained with PRLR isoform specific antibodies and analyzed. Scale bar indicates range of red scoring.

Article Snippet: After blocking in 5% normal goat serum (Jackson Laboratories, Bar Harbor, ME) prepared in PBS-0.1% Triton, slides were incubated with the PRLR isoform specific antibodies (10 μg/ml) for 2 hr at room temperature.

Techniques: Immunohistochemical staining, Staining

Journal: BMC Cancer

Article Title: Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

doi: 10.1186/1471-2407-10-678

Figure Lengend Snippet: Immunohistochemical analysis from lobular breast tissue arrays. Representative photographs of lobular carcinoma specimens to demonstrate absent, low, medium, or high amounts of fluorescent red intensity staining. 104 individual lobular specimens were stained with PRLR isoform specific antibodies and analyzed. Scale bar indicates range of red scoring.

Article Snippet: After blocking in 5% normal goat serum (Jackson Laboratories, Bar Harbor, ME) prepared in PBS-0.1% Triton, slides were incubated with the PRLR isoform specific antibodies (10 μg/ml) for 2 hr at room temperature.

Techniques: Immunohistochemical staining, Staining