Journal: International Journal of Proteomics

Article Title: Method for Recovery and Immunoaffinity Enrichment of Membrane Proteins Illustrated with Metastatic Ovarian Cancer Tissues

doi: 10.1155/2012/838630

Figure Lengend Snippet: No significant effect of the TD2 buffer was seen in either the rate of color development from the HRP conjugate or antigen affinity in commercial ELISA kits for (a) total human transferrin (Tf), (b and c) total human matrix metalloprotease 2 (MMP2) or 3 (MMP3), (d) total human V-akt murine thymoma viral oncogene homolog 1 (AKT1), or (e) soluble vascular endothelial growth factor receptor (sVEGFR). Apparent affinity constants (±one standard deviation) for separate serial dilutions for the kit standards are shown for each ELISA for both the recommended kit diluent and TD2 buffers. (Standard deviations are determined by partitioning the error of the estimate across the fitted parameters using the Jacobian matrix. In ELISA assays lacking experimental data defining the upper asymptotic limits of quantitation (e.g., VEGF R2 assay of ), this method produces very large errors when fitting the data with a nonlinear equation ).

Article Snippet: The effect of the diluted TD2 buffer on immunoassays was evaluated in several commercial ELISA kits, including human transferrin kit (Bethyl Laboratories, Montgomery, TX), Quantikine MMP-2 and MMP-3 (R&D Systems, Minneapolis, MN), and PathScan total p53 and PathScan total AKT1 (Cell Signaling, Danvers, MA).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Quantitation Assay

Journal: International Journal of Proteomics

Article Title: Method for Recovery and Immunoaffinity Enrichment of Membrane Proteins Illustrated with Metastatic Ovarian Cancer Tissues

doi: 10.1155/2012/838630

Figure Lengend Snippet: The concentrations of various protein biomarkers extracted from mixed metastatic ovarian tumors obtained from the omenta of various patients during surgical debulking as measured by ELISA. Proteins were extracted from the cryogenically ground tumor samples using a Barocycler with TD2 Buffer. The concentrations (± one standard deviation) of each biomarker are determined from the extract contained using the ELISA assays described in Figure 3 and extrapolated to that present in the tumor assuming 100% recovery. Transferrin was used as a ubiquitous control protein, which is indicative of the serum content of the sample. Some biomarkers were below the detection limits of the assay. MMP3 was below the limits of quantitation of the ELISA.

Article Snippet: The effect of the diluted TD2 buffer on immunoassays was evaluated in several commercial ELISA kits, including human transferrin kit (Bethyl Laboratories, Montgomery, TX), Quantikine MMP-2 and MMP-3 (R&D Systems, Minneapolis, MN), and PathScan total p53 and PathScan total AKT1 (Cell Signaling, Danvers, MA).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Biomarker Assay, Quantitation Assay, Concentration Assay

Journal: Biomolecules

Article Title: Effect of Exogenous Hydrogen Sulfide and Polysulfide Donors on Insulin Sensitivity of the Adipose Tissue

doi: 10.3390/biom12050646

Figure Lengend Snippet: Effect of insulin, Na 2 S, and Na 2 S 4 on phosphorylation of insulin receptor β-subunit ( a ), phosphorylation of IRS-1 ( b ), Akt1 phosphorylation at Ser 473 ( c ), and cAMP in the adipose tissue ( d ) in vivo. NaCl (control), Na 2 S (100 μmol/kg), or Na 2 S 4 (100 μmol/kg) were administered iv. 15 min before insulin (0.5 U/kg ip.). Samples of mesenteric adipose tissue were collected 30 min after insulin injection. ** p < 0.01, *** p < 0.001 vs. control group receiving only NaCl, # p < 0.05 vs. group receiving only insulin.

Article Snippet: We measured total Akt and Akt phosphorylated at Ser 473 using Cell Signaling Technology PathScan Total Akt1 Sandwich ELISA Kit (Kit #7170) and PathScan Phospho-Akt1 (Ser473) Sandwich ELISA Kit (#7160), respectively.

Techniques: In Vivo, Injection

Journal: Biomolecules

Article Title: Effect of Exogenous Hydrogen Sulfide and Polysulfide Donors on Insulin Sensitivity of the Adipose Tissue

doi: 10.3390/biom12050646

Figure Lengend Snippet: Effect of insulin, Na 2 S, and Na 2 S 4 on glucose uptake ( a ), glycerol release ( b ), tyrosine phosphorylation of IRβ ( c ), tyrosine phosphorylation of IRS-1 ( d ), Akt1 phosphorylation ( e ), and cAMP concentration ( f ) in mesenteric adipose tissue isolated from lean (white) and obese (black) rats. * p < 0.05, ** p < 0.0, *** p < 0.001 vs. samples not treated with insulin, Na 2 S, and Na 2 S 4 collected from the respective group of rats (lean or obese), ^ p < 0.05, ^^ p < 0.01, ^^^ p < 0.001 vs. adipose tissue samples treated in the same manner collected from lean rats, # p < 0.05 vs. samples from the respective group (lean or obese) treated with insulin alone.

Article Snippet: We measured total Akt and Akt phosphorylated at Ser 473 using Cell Signaling Technology PathScan Total Akt1 Sandwich ELISA Kit (Kit #7170) and PathScan Phospho-Akt1 (Ser473) Sandwich ELISA Kit (#7160), respectively.

Techniques: Concentration Assay, Isolation

Journal: Scientific Reports

Article Title: Resveratrol reverses the adverse effects of bevacizumab on cultured ARPE-19 cells

doi: 10.1038/s41598-017-12496-z

Figure Lengend Snippet: Effects of BEV + RES on Notch signaling. Bar graph depicts the relative mRNA expression levels of notch 4 and dll 4 in with respect to mRNA levels of gapdh in untreated ARPE-19 cells and those treated with BEV, RES and BEV + RES ( A ). Graphical representation of quantified mean fluorescent intensity of immunofluorescent images for NOTCH 4 and DLL 4 are shown ( B ). Representative immunofluorscent images showing positivity for NOTCH 4 and DLL4 in untreated and BEV, RES, BEV + RES treated ARPE-19 cells ( C ). Representative western blots of NOTCH 4, DLL4 and the downstream target HES 1 in untreated and treated ARPE-19 cells ( D ). Graphical representation of the densitometric analysis of western blot results with respect to the levels of GAPDH is shown ( E ). Graphical representation of PathScan ® ELISA results of untreated and cells treated with BEV, RES and BEV + RES ( F ). Representative Western blot showing phosphorylated and total MEK1/2 (i) and p44/42 MAPK (ii) ( G ). Statistical analysis was performed using student’s t-test (*P < 0.05,**P < 0.01, ***P < 0.005) (n ≥ 3). Scale bar = 5 µm.

Article Snippet: The quantity of phosporylated p-MEK1/2 (Ser217/221), p44/42MAPK(Thr202/Tyr204),p-SAPK/JNK(Thr183/Tyr185),p-Akt1(Ser473) and p-Akt1(Thr 308) were measured from cell lysates using human PathScan ® ELISA kits (Cell Signaling technology,Beverly,MA,USA).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay