Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 2. The mRNA levels of chemerin and its cognate receptors are altered in ob/ob mice. Relative mRNA levels of chemerin, CMKLR1, CCRL2, and GPR1 were determined in C57BL/6 and ob/ob white adipose, skeletal muscle, and liver tissue by quantitative real-time PCR. C57BL/6 expression served as the reference (expression 1.0) to which ob/ob mice were compared (n 5–10). Each bar represents the mean SEM. *, P 0.05, unpaired t test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 1. Chemerin and cognate receptors are differentially expressed in tissues important in glucose homeostasis. Relative mRNA levels of chemerin, CMKLR1, CCRL2, and GPR1 were determined in C57BL/6 mouse white adipose, skeletal muscle, and liver tissues by quantitative real-time PCR. White adipose served as the reference tissue (expression 1.0) to which all other tissues were compared. n 4–5. Each bar represents the mean SEM. *, P 0.05, one-way ANOVA followed by Bonferroni’s multiple-comparison test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Comparison

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 3. The mRNA levels of chemerin and its cognate receptors are altered in db/db mice. Relative mRNA levels of chemerin, CMKLR1, CCRL2, and GPR1 were determined in C57BL/6 and db/db white adipose, skeletal muscle, and liver tissue by quantitative real-time PCR. C57BL/6 expression served as the reference (expression 1.0) to which ob/ob mice were compared (n 5–10). Each bar represents the mean SEM. *, P 0.05, unpaired t test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 4. Serum chemerin levels are elevated in mouse models of obesity and diabetes. Blood was collected using cardiac puncture and was allowed to coagulate for 2 h. A and B, The resulting serum was analyzed for total chemerin using a mouse chemerin ELISA (A) and bioactive chemerin levels using a CMKLR1 Tango assay (B). C, We then calculated the ratio of bioactive to total chemerin levels. n 5. Each bar is the mean SEM. **, P 0.01 vs. C57BL/6, one-way ANOVA followed by Bonferroni’s multiple-comparison test. D, Total chemerin protein was also examined in white adipose (WA), liver (LV), and skeletal muscle (SM) tissues using Western blotting. ND, Not detected. Values represent mean relative densitometry data SEM. *, P 0.05, unpaired t test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

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

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 5. Chemerin treatment exacerbates glucose intolerance in ob/ob mice. Vehicle or chemerin (4 or 40 ng/g) was injected ip to C57BL/6 (A and C) and ob/ob (B and D) mice with glucose (2 mg/g). A and B, Blood samples were collected over a 2-h period, and blood glucose levels were analyzed. C and D, Serum samples were also collected throughout the GTT and analyzed for serum insulin levels (n 10). Each bar represents mean SEM. *, P 0.05; **, P 0.01 vs. vehicle, two-way ANOVA followed by Bonferroni’s multiple-comparison test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: Injection, Comparison

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 7. Chemerin treatment exacerbates glucose intolerance in DIO mice. Vehicle or chemerin (4 or 40 ng/g) was injected ip to C57BL/6 (A and C) and DIO (B and D) mice with glucose (2 mg/g). A and B, Blood samples were collected over a 2-h period, and blood glucose levels were analyzed. C and D, Serum samples were also collected throughout the GTT and were analyzed for serum insulin levels (n 10). Each bar represents mean SEM. *, P 0.05 vs. vehicle, two-way ANOVA followed by Bonferroni’s multiple-comparison test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: Injection, Comparison

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 6. Chemerin treatment exacerbates glucose intolerance in db/db mice. Vehicle or chemerin (4 or 40 ng/g) was injected ip to C57BL/6 (A and C) and db/db (B and D) mice with glucose (2 mg/g). A and B, Blood samples were collected over a 2-h period, and blood glucose levels were analyzed. C and D, Serum samples were also collected throughout the GTT and were analyzed for serum insulin levels. n 10. Each bar represents mean SEM. *, P 0.05; **, P 0.01 vs. vehicle, two-way ANOVA followed by Bonferroni’s multiple- comparison test.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: Injection, Comparison

Journal: Endocrinology

Article Title: Chemerin exacerbates glucose intolerance in mouse models of obesity and diabetes.

doi: 10.1210/en.2009-1098

Figure Lengend Snippet: FIG. 8. Chemerin decreases in vivo tissue glucose uptake in db/db mice. Vehicle or chemerin (4 or 40 ng/g) was injected ip to C57BL/6 (A) and db/db (B) mice with glucose (2 mg/g) and 10 Ci 2-DOG. Blood samples were collected over a 1-h period, and blood glucose concentration and radioactivity were measured. After 60 min, mice were euthanized, tissues were snap frozen and homogenized, and radioactivity was measured. GSA was calculated by dividing blood radioactivity by blood glucose concentration and calculating the area under the curve. Tissue radioactivity disintegrations per minute (DPM) was normalized to GSA and the mass of tissue homogenized (n 5–10). Each bar represents the mean SEM. *, P 0.05, unpaired t test. SKM, Skeletal muscle; WA, white adipose.

Article Snippet: After this, the membranes were incubated overnight at 4 C with an antimouse chemerin antibody (AF2325; R&D Systems, Minneapolis, MN) diluted 1:500 in blocking solution.Afterwashingfourtimesfor5mininTBST,themembranewas further incubated with a horseradish peroxidase-conjugated donkey antigoat IgG secondary antibody (1:10,000 in blocking buffer) for 1 h at room temperature.

Techniques: In Vivo, Injection, Concentration Assay, Radioactivity

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 2. a) Surface-enhanced Raman scattering (SERS) spectra of chemerin_pooled follicular fluid (FF) samples and b) score plot of principal component (PC) of polycystic ovary syndrome (PCOS) and non-PCOS data of the first sample group.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques:

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 3. Surface-enhanced Raman scattering (SERS) spectra of chemerin in follicular fluid (FF) a) non-polycystic ovary syndrome (PCOS) and b) PCOS samples.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques:

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Figure 5 shows an increase in the peak intensity in the Raman spectrum with increased chemerin concentration. Fifty spectra of the third group of known samples (ten Raman spectra collected to ensure consistency of the replicated measurements) were used to build a PLS model, and twenty spectra of the second group were used to predict the chemerin concentration using the model. A TSV model was then developed using 67% of the known samples as a calibration set, and 33% as a validation set. These were chosen randomly by the Unscramble software.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques: Concentration Assay, Biomarker Discovery, Software

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 6. Loading plot (a) and score plot (b) of principal component (PC) of chemerin in phosphate-buffered saline (PBS) samples.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques: Saline

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 5. Surface-enhanced Raman scattering (SERS) spectra of chemerin in phosphate-buffered saline (PBS) samples.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques: Saline

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 8. Partial least squares (PLS) regression model for chemerin concentration prediction in 650 to 1450 cm−1 spectral range using test set validation.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques: Concentration Assay, Biomarker Discovery

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 9. Baseline corrected surface-enhanced Raman scattering (SERS) spectra of a) polycystic ovary syndrome (PCOS) and b) non-PCOS patient samples spiked with chemerin.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques:

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 13. Raman spectra of chemerin in phosphate-buffered saline (PBS).

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques: Saline

Journal: Biomedical Optics Express

Article Title: Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome

doi: 10.1364/boe.9.000801

Figure Lengend Snippet: Fig. 14. The loading plots of higher principal components (PCs) of chemerin in phosphate- buffered saline (PBS) samples.

Article Snippet: Non-specific binding was blocked using 5% skim milk in TBS-T (0.3%; 1 h, RT), and then incubated (overnight, 4°C) with diluted goat anti-human chemerin (1: 1000; R&D System #AF2324) in TBS-T (0.05%).

Techniques: Saline

Journal: Endocrinology

Article Title: Chemerin, a novel regulator of follicular steroidogenesis and its potential involvement in polycystic ovarian syndrome.

doi: 10.1210/en.2012-1424

Figure Lengend Snippet: FIG. 2. Chemerin levels in serum and ovarian cells in CTL and DHT-treated rats. A, Serum (0.5 l) from CTL and DHT-treated rats was subjected to Western blot and immunoblotted with antichemerin antibody. Immunoblots shown are six representative serum samples in each experimental group, whereas each histogram shows results from 12 rats. B, Chemerin and CMKLR1 contents in whole-ovary extracts from CTL and DHT-treated rats were analyzed by Western blot and normalized to GAPDH. Each sample was collected from one ovary of individual CTL or DHT rats (15 rats per histogram), whereas six representative samples were presented in the immunoblots. C, Total RNA were extracted from whole ovaries of CTL and DHT-treated rats. Each sample was collected from one ovary of CTL or DHT rats (nine rats per histogram). The mRNA abundances of chemerin and CMKLR1 were analyzed by real-time PCR and normalized to the expression of GAPDH. D, Granulosa cells were collected from four to five rats of the CTL or DHT group and pooled for total RNA extraction. The mRNA abundances of chemerin and CMKLR1 were analyzed by real-time PCR and normalized to the expression of GAPDH. Data are presented as mean SEM of five independent experiments. Panels A–D used an unpaired Student t test. *, P 0.05, ***, P 0.001 compared with CTL group. NS, Not significant.

Article Snippet: Recombinant mouse active chemerin and antichemerin antibody were from R&D Systems (Minneapolis, MN).

Techniques: Western Blot, Real-time Polymerase Chain Reaction, Expressing, RNA Extraction

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Chemerin in primary human hepatocytes (PHH) and hepatocyte cell lines. ( A ) Chemerin mRNA in HepG2, Huh7 and Hep3B cells and PHH (n = 3–6); ( B ) and ( C ) Chemerin protein in HepG2, Hep3B, PLC/PRF/5 (PLC), and Huh7 cells and PHH; ( D ) Chemerin in supernatants of HepG2, Huh7, Hep3B and PLC/PRF/5 (PLC) cells and PHH (n = 7–8); ( E ) Chemerin in HepG2 cells transfected with scrambled (scr) or chemerin (chem) siRNA; ( F ) Chemerin in PHH transfected with scrambled or chemerin siRNA; ( G ) Chemerin in the supernatant of PHH transfected with scrambled or chemerin siRNA; ( H ) Chemerin in the supernatant of PHH transfected with scrambled or chemerin siRNA measured by ELISA (n = 6 different donors). * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical test used: unpaired and paired Student´s t-test.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Transfection, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Chemerin in non-tumor (NT) and tumor tissue (TT) of 21 HCC patients with non-viral disease etiology. ( A ) Chemerin mRNA, the number in the figure indicates the p-value; ( B ) Chemerin protein in NT and TT of three different patients (P1 to P3); ( C ) Quantification of chemerin protein which was analyzed by immunoblot in 21 patients. GAPDH was used for normalization; ( D ) Ratio of TT to NT chemerin protein in patients with cryptic disease etiology (n = 11) or NAFLD (n = 10); ( E ) Chemerin in tumors of patients with grade 1 (well differentiated tumor) or grade 2 (moderately differentiated tumor); ( F ) Chemerin protein in the tumors of 20 patients stratified for TNM classification. * p < 0.05; ** p < 0.01. Statistical test used: Mann-Whitney U test and one-way ANOVA with posthoc Bonferroni.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Western Blot, MANN-WHITNEY

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Post hoc Power Analysis using G*Power 3.1.6. T Test-means: Wilcoxon signed-rank test (matched pairs). Output parameters of the analysis are shown for the different groups.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques:

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Chemerin in non-tumor (NT) and tumor tissue (TT) of 24 HCC patients with viral disease etiology. ( A ) Chemerin protein of three different patients. ( B ) Quantification of chemerin protein in 10 HBV, 11 HCV and 3 HBV/HCV infected patients. Coomassie stained membrane was used for normalization. ( C ) Ratio of TT to NT chemerin protein in 15 patients without and eight patients with fatty liver (FL). ( D ) Chemerin protein stratified for TNM classification in 23 patients. ( E ) Chemerin protein stratified for TNM classification in HBV infected patients (there was one patient with pT3a, and statistical test was not possible for this comparison). n.s. not significant. ** p < 0.01. Statistical test used: Mann-Whitney U test and one-way ANOVA with posthoc Bonferroni.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Infection, Staining, Membrane, Comparison, MANN-WHITNEY

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Analysis of chemerin variants by immunoblot. ( A ) Human chemerin isoforms 163, 157, 156 and 155 were overexpressed in HepG2 cells and protein was detected by three different antibodies (C, control-transfected cells). ( B ) Quantification of chemerin protein (n = 2).

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Western Blot, Control, Transfection

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Chemerin protein in non-tumor (NT) and tumor tissue (TT) of HCC patients. Chemerin protein in NT and TT of six patients (P1 to P6) was analyzed with three different antibodies. pAB, polyclonal antibody; mAB monoclonal antibody; SE, short exposure time of the membrane.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Membrane

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Effect of retinoic acid and tazarotene on chemerin and CMKLR1. ( a ) HepG2 cells were incubated with increasing concentrations of retinoic acid for 24 h and LDH was measured in the supernatants (n = 4); ( b ) HepG2 cells were incubated with increasing concentrations of tazarotene for 24 h and LDH was measured in the supernatants (n = 4); ( c ) HepG2 cells were incubated with retinoic acid (1 and 5 µM) for 24 h and chemerin was measured in the supernatants (n = 4); ( d ) HepG2 cells were incubated with tazarotene (0.1 and 0.5 µM) for 24 h and chemerin was measured in the supernatants (n = 4); ( e ) HepG2 cells were incubated with retinoic acid (1 and 5 µM) for 24 h and CMKLR1 was analyzed by immunoblot (n = 2); ( f ) HepG2 cells were incubated with tazarotene (0.1 and 0.5 µM) for 24 h and CMKLR1 was analyzed by immunoblot (n = 2). Bovine serum albumin (BSA) dissolved in EtOH and dimethylsulfoxid (DMSO) served as solvent controls. * p < 0.05, *** p < 0.001. Statistical test used: Paired t-test.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Incubation, Western Blot, Solvent

Journal:

Article Title: Chemerin, a Novel Adipokine in the Regulation of Angiogenesis

doi: 10.1210/jc.2010-0042

Figure Lengend Snippet: Genes within the genome that was associated with plasma chemerin levels

Article Snippet: Chemerin ELISA Chemerin levels in 1354 human plasma samples from the SAFHS were determined using a sandwich ELISA developed with commercially available unlabeled and biotinylated polyclonal antihuman chemerin antibodies (R&D Systems, Minneapolis, MN) as previously described ( 11 ).

Techniques: Clinical Proteomics

Journal:

Article Title: Chemerin, a Novel Adipokine in the Regulation of Angiogenesis

doi: 10.1210/jc.2010-0042

Figure Lengend Snippet: Effect of recombinant chemerin concentration on blood vessel formation by endothelial cells in a coculture system. A coculture of endothelial cells and fibroblasts was used to study the tube formation by endothelial cells. The AngioKit was seeded with cells on d 0, and the optimized growth medium was changed on d 1, 3, 5, and 7. The cells were then fixed and stained for CD31 on d 9. Suramin (20 μm) and VEGF (2 ng/ml) were used as negative and positive controls, respectively. Recombinant human chemerin was added to each well at 0.1, 0.3, 1, 3, and 10 ng/ml. A, Control: cell medium only. B, Inhibition of tubule formation to control level by suramin (20 μm). C, VEGF (2 ng/ml) induced tubule formation as well as VEGF-induced increased branching of tubules and formation of in vitro capillaries. D–F, Coculture of endothelial cells and fibroblasts (AngioKit) after 9 d incubation in optimized growth media supplemented with 0.1, 1, and 10 ng/ml of recombinant human chemerin, respectively. The number of microtubules (G), tubule anastomoses (H), and microtubule length (I) were measured using an AngioSys image analysis system. Four images were captured per well from four independent quadrants. Experiments were run in triplicate, and data were expressed as the mean ± sem. *, P < 0.05, **, P < 0.01 compared with the vehicle control.

Article Snippet: Chemerin ELISA Chemerin levels in 1354 human plasma samples from the SAFHS were determined using a sandwich ELISA developed with commercially available unlabeled and biotinylated polyclonal antihuman chemerin antibodies (R&D Systems, Minneapolis, MN) as previously described ( 11 ).

Techniques: Recombinant, Concentration Assay, Staining, Control, Inhibition, In Vitro, Incubation

Journal:

Article Title: Chemerin, a Novel Adipokine in the Regulation of Angiogenesis

doi: 10.1210/jc.2010-0042

Figure Lengend Snippet: The effect of PD, a MEK1 inhibitor, on the angiogenic properties of chemerin in endothelial cells in a coculture system. The coculture of endothelial cells and fibroblasts (AngioKit) was used to study the formation of new blood vessels. The AngioKit was seeded with cells on d 0, and the medium was changed on d 1, 3, 5, and 7. Recombinant human chemerin was added at 3 and 10 ng/ml, respectively (in the presence or absence of 25 μm PD 98059). Cells were then fixed, and stained for CD31 on d 9. A, Control: cell medium only. B, Growth media with 0.1% DMSO. C, VEGF-positive control (2 ng/ ml). D, PD (25 μm). E, Recombinant human chemerin (3 ng/ml). F, Recombinant human chemerin (3 ng/ml) + PD (25 μm). The number of microtubule junctions (G), total number of microtubules (H), total microtubule length (I), and total tubule area (J) were measured using AngioSys image analysis system. Four images were captured per well from four independent quadrants. Experiments were run in triplicate, and data were expressed as the mean ± sem. *, P < 0.05, **, P < 0.01 compared with the vehicle control.

Article Snippet: Chemerin ELISA Chemerin levels in 1354 human plasma samples from the SAFHS were determined using a sandwich ELISA developed with commercially available unlabeled and biotinylated polyclonal antihuman chemerin antibodies (R&D Systems, Minneapolis, MN) as previously described ( 11 ).

Techniques: Recombinant, Staining, Control, Positive Control

Journal: Cancers

Article Title: Chemerin Is Induced in Non-Alcoholic Fatty Liver Disease and Hepatitis B-Related Hepatocellular Carcinoma

doi: 10.3390/cancers12102967

Figure Lengend Snippet: Chemerin protein in non-tumor (NT) and tumor tissue (TT) of HCC patients. Chemerin protein in NT and TT of six patients (P1 to P6) was analyzed with three different antibodies. pAB, polyclonal antibody; mAB monoclonal antibody; SE, short exposure time of the membrane.

Article Snippet: A polyclonal (AF2324, RRID: AB_416577) and a monoclonal chemerin antibody (MAB2324, RRID: AB_2175697) were from R&D Systems (Wiesbaden, Germany).

Techniques: Membrane