anti gapdh v 18  (Santa Cruz Biotechnology)

Journal: International journal of molecular sciences

Article Title: Establishment of a Human iPSC Line from Mucolipidosis Type II That Expresses the Key Markers of the Disease.

doi: 10.3390/ijms26083871

Figure Lengend Snippet: Figure 3. Endogenous expression of pluripotency markers in ML II iPSCs. (A) Expression levels of the pluripotency genes NANOG, OCT4, and SOX2 in ML II iPSCs relative to the parental ML II fibroblasts, measured by RT-qPCR. The results were normalized to the housekeeping gene GAPDH, and expression levels were calculated relative to the expression in the fibroblasts by the 2−∆∆Ct

Article Snippet: Total protein loaded was controlled by incubation with a monoclonal anti-GAPDH (V-18) antibody (sc-20357, Santa Cruz Biotechnology, Dallas, Texas, EUA) and the secondary antibody donkey anti-goat IgG-HRP (sc-2020, Santa Cruz Biotechnology, Dallas, Texas, EUA).

Techniques: Expressing, Quantitative RT-PCR

Journal: International journal of molecular sciences

Article Title: Establishment of a Human iPSC Line from Mucolipidosis Type II That Expresses the Key Markers of the Disease.

doi: 10.3390/ijms26083871

Figure Lengend Snippet: Figure 8. Western blotting for alpha-galactosidase. (A) Cellular extracts from WT and ML II fibroblasts and iPSCs. GAPDH was used as a loading control, and densitometric analyses of alpha-galactosidase levels were normalized for GAPDH. Two independent experiments were performed. (B) Conditioned media from WT and ML II iPSCs analyzed for alpha-galactosidase.

Article Snippet: Total protein loaded was controlled by incubation with a monoclonal anti-GAPDH (V-18) antibody (sc-20357, Santa Cruz Biotechnology, Dallas, Texas, EUA) and the secondary antibody donkey anti-goat IgG-HRP (sc-2020, Santa Cruz Biotechnology, Dallas, Texas, EUA).

Techniques: Western Blot, Control

Journal: Molecular Cancer

Article Title: Runx2 transcriptome of prostate cancer cells: insights into invasiveness and bone metastasis

doi: 10.1186/1476-4598-9-258

Figure Lengend Snippet: Establishment of C4-2B/Rx2 dox sub-line with conditional Runx2 expression . A, Schematic diagram of the pSLIK-based lentiviral vector. Initially developed for expression of shRNAs the pSLIK vector was used in the present study to express Runx2. The Hygromycin resistance marker ( Hyg ) and the Dox-dependent activator protein rtTA3 are constitutively expressed under the control of the Ubi-c promoter. Upon treatment with Dox, rtTA binds to its tetracycline responsive elements ( TRE ) and drives expression of the inserted cDNA (Flag-Runx2). The Runx2 block diagram depicts its glutamine/alanine-rich QA domain, the DNA-binding Runt domain, and the proline/serine/threonine-rich PST domain. Arrowhead indicates the position of the R265D and R268D mutations in Runx2-M, which eliminate Runx2's DNA binding function. B, Whole cell extracts, prepared from C4-2B/Rx2 dox and C4-2B/Rx2-M dox cells treated with the indicated concentrations of Dox, were subjected to western blot analysis using anti-Flag antibodies. C, Total RNA was extracted from C4-2B/Rx2 dox cells treated with Dox or vehicle, as well as from PC3 high cells, and the mRNA levels of Runx2 (and GAPDH as control) were measured by RT-qPCR. D, Whole cell extracts were prepared from C4-2B/Rx2 dox cells treated with Dox as indicated and from ROS 17.8/2 osteoblastic cells , and subjected to western blot analysis using anti-Runx2 antibodies. The same blot was re-probed with anti-Tubulin antibodies as loading control. E, C4-2B/Rx2 dox and C4-2B/Rx2-M dox cells were transiently transfected with the 6XOSE2-luciferase reporter plasmid and subjected to luciferase assay. Dotted line represents the background luciferase activity with no cell extract. F, C4-2B/Rx2 dox cells were treated with Dox and levels of the indicated transcripts were measured by RT-qPCR and corrected for that of GAPDH. In Figure 1, bars represent Mean ± SEM (n = 3) from a representative experiment, which was repeated at least three times with similar results. Abbreviations used: Dox, Doxycycline; Veh, vehicle; BSP, Bone Sialoprotein; MMP9, Matrix Metalloprotein-9; OC, Osteocalcin.

Article Snippet: The anti-PIP antibody (ab 62363) was purchased from abcam Inc., Cambridge, MA; and anti-GAPDH (V-18), anti-TGFß-RIII (Sc 28975), and anti-SDF-1 (Sc-28876) antibodies were purchased from Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA.

Techniques: Expressing, Plasmid Preparation, Marker, Blocking Assay, Binding Assay, Western Blot, Quantitative RT-PCR, Transfection, Luciferase, Activity Assay