Journal: The Journal of clinical endocrinology and metabolism

Article Title: Effect of a vitamin D3 analogue on keratinocyte growth factor-induced cell proliferation in benign prostate hyperplasia.

doi: 10.1210/jcem.85.7.6690

Figure Lengend Snippet: FIG. 4. Western blot analysis of total lysates from BPH cells probed for tyrosine-phosphorylated proteins (A) and for KGF-R (B). Molecular weight markers are indicated to the right of each blot. A, Total extracts (25 mg) from BPH cells untreated (control) or treated for 5 min with analogue (V) (10 nmol/L), KGF (10 ng/mL), or a combination of analogue (V) (10 nmol/L) and KGF (10 ng/mL), were separated by 8% SDS-PAGE and transferred onto nitrocellulose membranes and then probed with PY20-HRP antibody (1:1000). After treatment with KGF, there is an evident increase in tyrosine-phosphorylation of a 120-kDa protein. Simultaneous incubation with analogue (V) completely prevented such an increase, whereas analogue (V) alone was without effect. B, After stripping, the same blot as in A was probed for KGF-R. Western blot analysis with an anti-KGF-R antibody (1 mg/mL) reveals the same 120-kDa protein band as in A. The blot is representative of three separate experiments.

Article Snippet: Horse radish peroxidase-conjugated monoclonal antiphosphotyrosine antibody (PY20-HRP) was purchased from ICN Biomedicals, Inc. (Costa Mesa, CA) and anti-KGF receptor (anti-KGF-R) antibody from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Western Blot, Molecular Weight, Control, SDS Page, Phospho-proteomics, Incubation, Stripping Membranes