human sdf1a elisa kits  (R&D Systems)

Journal: Cancer Research

Article Title: Altered TGF-β Signaling in a Subpopulation of Human Stromal Cells Promotes Prostatic Carcinogenesis

doi: 10.1158/0008-5472.can-10-3142

Figure Lengend Snippet: Figure 4. Molecular consequences of heterogeneous stromal cell mixes. BHPrS1EV, BHPrS1DN, and BHPrS1EV/BHPrS1DN mixtures were cultured for 48 hours before RNA and protein isolation. A, RT-PCR analysis showed increased SDF1a expression and down-regulation of TGFbR3 in BHPrS1EV/BHPrS1DN compared with the other groups. B, PCR array analysis validated the increased expression of TGF-b1 in BHPrS1EV/BHPrS1DN

Article Snippet: Quantification of TGF-b1 and SDF1a in the CM were assessed by ELISA according to the manufacturer's protocol [human TGF-b1 (DB100B) and human SDF1a (DSA00) Quantikine from R&D Systems, Inc, Minneapolis, MN].

Techniques: Cell Culture, Isolation, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Cancer Research

Article Title: Altered TGF-β Signaling in a Subpopulation of Human Stromal Cells Promotes Prostatic Carcinogenesis

doi: 10.1158/0008-5472.can-10-3142

Figure Lengend Snippet: Figure 5. Induction of CAF phenotype by overexpression of TGF-b1 and SDF1a in normal prostate fibroblasts. A, ELISA was used to test and corroborate the expression of the transgenes in fibroblasts. Fibroblasts expressing constitutively active TGF-b1 ligand showed an increased SDF1a expression. However, overexpressing SDF1a in fibroblasts had no effect on TGF-b1 levels. B, BHPrS1TGF-b1 and BHPrS1SDF1a were recombined with BPH1 cells and xenografted under the kidney capsule for about 8 weeks. The volume of the tumors composed of BHPrS1TGF-b1þBPH1 and BHPrS1SDF1aþBPH1 were significantly larger than the controls. Note the invasive characteristic of the tumors (arrows) C. Histological examination revealed malignant trasnformation of the epithelial cells (i–iii). TGF-b1-expressing fibroblasts had a greater impact on the proliferation of BPH1 cells as shown by Ki67 staining as compared with BHPrS1SDF1a cells, (iv–vi). BHPrS1EV fibroblasts exhibited light blue stromal Trichrome staining indicative of some collagen in the stroma. In contrast, recombinants composed of BHPrS1SDF1a stained predominantly red suggesting a more muscular phenotype. Intense blue Trichrome staining in the stroma of BHPrS1TGF-b1þBPH1 recombinants revealed the extensive collagen deposition in these tumors (vii–ix). Intense SDF1a staining corroborated the secretion of the chemokine in grafts composed of SDF1a-expressing compared with TGF-b1-expressing fibroblasts (x–xii).

Article Snippet: Quantification of TGF-b1 and SDF1a in the CM were assessed by ELISA according to the manufacturer's protocol [human TGF-b1 (DB100B) and human SDF1a (DSA00) Quantikine from R&D Systems, Inc, Minneapolis, MN].

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Expressing, Staining