igfbp3 elisa kit  (Elabscience Biotechnology)

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: EGFRvIII and TGF-β synergistically promoted expression of IGFBP3 (A) qRT-PCR analysis of IGFBPs mRNA in U87-vector and-EGFRvIII with TGF-β (10 ng/mL) treated for 12 h and 24 h. (B) Western blotting analysis of expression of IGFBP3 in U87-vector and-EGFRvIII cells treated with TGF-β for indicated times, β-actin was used as a loading control. (C) The expression of IGFBP3 in cell culture supernatant was detected by ELISA and western blotting under TGF-β treatment. (D) Western blotting analysis of the expression of total and phosphorylation of EGFRvIII, Smad2/3, ERK1/2, AKT in U87-EGFRvIII cells or U87-vector cells treated with TGF-β. (E) The analysis of immunoprecipitation (IP) showed the firm association of EGFRvIII with TGF-β in U87-EGFRvIII cell. (F) Western blotting analysis of the expression level of IGFBP3, p-EGFRvIII, and downstream signal proteins in U87-EGFRvIII cells treated with the TGF-βR inhibitor LY2157299 (0, 5, 10 and 20 μM) and then stimulated with TGF-β. (G) Protein expression levels of total and phosphorylated of EGFRvIII, ERK1/2, AKT and IGFBP3 in U87-EGFRvIII cells pretreated with U0126 (15 μM) or LY294002 (15 μM) and then stimulated cells with TGF-β. Values are the means ± SD of three independent experiments. ∗p<0.05; ∗∗p<0.01; ∗∗∗p<0.001. See also Figure S1 .

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques: Expressing, Quantitative RT-PCR, Plasmid Preparation, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Immunoprecipitation

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: TGF-β induced IGFBP3 and EGFRvIII nuclear translocation (A)Immunofluorescence staining of EGFRvIII (green), IGFBP3 (red) and DAPI (blue) in U87-EGFRvIII cells after TGF-β treatment. Scale bar = 25 μm, arrows showed changes in the co-localization of EGFRvIII and IGFBP3. (B) Western blotting analysis of the nuclear protein levels of IGFBP3, total and phosphorylated of EGFRvIII, Smad2/3 and ERK1/2 after TGF-β treatment. (C) The expression level of IGFBP3, TGF-β and EGFRvIII downstream signal proteins were detected in the nuclear and cytoplasmic of U87-EGFRvIII cells treated with LMB and then stimulated with TGF-β. (D) ELISA analysis of the expression level of secreted IGFBP3 in U87-EGFRvIII cell culture supernatant pretreated with LMB and then stimulated with TGF-β. (E) The protein expression level of IGFBP3 in cell supernatant of U87-EGFRvIII cells pretreated with LMB and then stimulated with TGF-β. ∗∗∗p<0.001. See also Figure S2 .

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques: Translocation Assay, Immunofluorescence, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: TGF-β regulates the transcription of IGFBP3 by activating the transcription factor c-Jun (A) The mRNA of c-Jun was detected in U87-EGFRvIII by qRT-PCR after TGF-β treatment. (B) Western blotting analysis of c-Jun protein levels in U87-vector or U87-EGFRvIII cells treated with TGF-β for indicated times. (C) Western blotting analysis of the expression level of c-Jun in U87-EGFRvIII cells treated with LY2157299 (0, 5, 10 and 20 μM) and then stimulated with TGF-β. (D) Protein expression levels of c-Jun in U87-EGFRvIII cells pretreated with U0126 and LY294002 and then stimulated cells with TGF-β. (E) Schematic diagram depicting the regulatory sequences of the IGFBP3 promoter region. (F) Luciferase assays for specific binding of IGFBP3 promoter (WT) or (mutant)-Luc plasmid to c-Jun plasmid transfected with U87-EGFRvIII. (G) U87-EGFRvIII cells were transfected with Scramble (Scr) or c-Jun specific siRNA, then treated with TGF-β for 24 h, the expression level of c-Jun and IGFBP3 mRNA were determined by qRT-PCR analysis. (H) Western blotting analysis of the effect of c-Jun knockdown on TGF-β-induced upregulation of c-Jun, IGFBP3 and EGFRvIII proteins level. ∗p<0.05; ∗∗p<0.01; ∗∗∗p < 0.001. See also Figure S3 .

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

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

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: IGFBP3 enhanced the malignant behavior of EGFRvIII-expressing cells under the administration of exogenous TGF-β (A) U87-vector and U87-EGFRvIII cells were transfected with Scr or IGFBP3 specific siRNA, and the expression level of IGFBP3 protein were determined by western blotting. (B) Effect of IGFBP3 knockdown on cell cycle distribution under the treatment of TGF-β. (C) Western blotting analysis the effect of IGFBP3 knockdown on S-phase related protein expression under the treatment of TGF-β. (D) Knockdown of IGFBP3 decreases the migration and invasion capacities induced by TGF-β in U87-EGFRvIII cell, quantitative images of migrant and invasive cells were shown based on wound healing and transwell assay. (E) Western blotting analysis of the effect of IGFBP3 knockdown on TGF-β-induced upregulation of p-EGFRvIII, p-Smad2/3, p-ERK1/2, p-AKT and MMP2/9 proteins level in U87-EGFRvIII cell and U87-vector cell. (F) Anti-IGFBP3 decreases the invasion capacities induced by TGF-β. (G) Western blotting analysis of the effect of anti-IGFBP3 on TGF-β-induced upregulation of p-EGFRvIII, p-Smad2 and p-AKT proteins level in U87-EGFRvIII cell. ∗p<0.05; ∗∗p<0.01; ∗∗∗p<0.001. See also Figure S5 .

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques: Expressing, Plasmid Preparation, Transfection, Western Blot, Migration, Transwell Assay

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: Effect of IGFBP3 knockdown in vivo growth of U87-EGFRvIII cells using mice subcutaneous xenograft models. Nude mice were subcutaneously injected with pretreated U87-EGFRvIII- siIGFBP3 and U87-EGFRvIII- siControl cells (n = 5 for each group) (A) Tumor growth curves show tumor size measured every two days. (B) Tumor images and weights of the removed subcutaneous tumors after inoculation for 21 days. (C) The overall survival of mice in the U87-EGFRvIII-shControl and U87-EGFRvIII-shIGFBP3 cells implanted groups. Median survival of each group was 28 days for shControl, 38 days for shIGFBP3. n = 10, ∗∗p < 0.01 compared to the control, logrank test. (D) Representative images of IGFBP3, Ki-67 and tumor angiogenesis stained with CD31 in mouse subcutaneous tumors by IHC. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques: In Vivo, Injection, Staining

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: EGFRvIII and IGFBP3 expression levels in glioma tissues (A) Representative IHC stained images of glioma tissues using the anti-EGFRvIII and anti-IGFBP3 antibodies. (B) EGFRvIII and IGFBP3 protein expression in tissue specimens from Normal tissue and WHO I ∼ IV glioma patients. (C) Correlation analysis of EGFRvIII and IGFBP3 in tissues. ∗p < 0.05; ∗∗p < 0.01.

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques: Expressing, Staining

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet: Schematics illustration depicting the key findings of this study TGF-β and EGFRvIII transactivation induced the positive feedback loop of IGFBP3 to promote the progression of EGFRVII-positive GBMs, and targeted IGFBP3 can correct this vicious cycle to combat EGFRvIII-positive GBMs.

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques:

Journal: iScience

Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

doi: 10.1016/j.isci.2023.106639

Figure Lengend Snippet:

Article Snippet: Human IGFBP3 ELISA kits , Raybiotech , Cat#ELH-IGFBP3.

Techniques: Luciferase, Microarray, Recombinant, Lysis, Transfection, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Reporter Gene Assay, BIA-KA, Cell Cycle Assay, Sequencing, shRNA, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software

Journal: Journal of Molecular Endocrinology

Article Title: CTCF variant begets to short stature by down-regulation of IGF1

doi: 10.1530/JME-22-0193

Figure Lengend Snippet: Genetic information and response to growth hormone therapy in the patient.

Article Snippet: The protein expression levels of IGF1 and IGFBP3 were quantified using Human IGF1 ELISA Kit (Elabscience, Wuhan, China) and Human IGFBP-3 ELISA Kit (Elabscience), respectively.

Techniques:

Journal: Journal of Molecular Endocrinology

Article Title: CTCF variant begets to short stature by down-regulation of IGF1

doi: 10.1530/JME-22-0193

Figure Lengend Snippet: The CTCF R567W variant inhibition of IGFs gene expression. (A) qPCR analysis of the expression level of CTCF mRNA in the wild and mutant groups in three different cell lines. (B) Western blot analysis of the expression level of CTCF protein in the wildtype and mutant groups in LO2 cells. FLAG is a label protein of vectors. (C) In HEK-293T, LO2, and LCLs, mRNA levels of IGF1 were measured using quantitative PCR. (D) IGF1 protein levels were analyzed with ELISA. (E) In HEK-293T, LO2, and LCLs, mRNA levels of IGFBP3 were measured using quantitative PCR. (D) IGFBP3 protein levels were analyzed with ELISA. For quantitative PCR, results were normalized to β-actin as a reference and compared with control cells transfected with an empty vector plasmid ( n ≥ 3; mean ± s.e.m. shown; *** P < 0.001 by Student t -test). For Western blotting, results were normalized to β-actin as a reference ( n = 3; mean ± s.e.m. shown; * P < 0.05, ** P < 0.01, and *** P < 0.001). A full color version of this figure is available at https://doi.org/10.1530/JME-22-0193 .

Article Snippet: The protein expression levels of IGF1 and IGFBP3 were quantified using Human IGF1 ELISA Kit (Elabscience, Wuhan, China) and Human IGFBP-3 ELISA Kit (Elabscience), respectively.

Techniques: Variant Assay, Inhibition, Gene Expression, Expressing, Mutagenesis, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Transfection, Plasmid Preparation

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Clinical characteristics of samples in two cohorts

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques: Biomarker Discovery

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Level and ROC curve analysis of serum IGFBP3 for EJA. Serum IGFBP3 levels in normal controls, EJA and early-stage EJA patients in training cohort ( A ) and validation cohort ( B ) are shown in scatter plot and box plot. The lines in the box are means. The ROC curves of serum IGFBP3 to distinguish EJA and early-stage EJA patients and normal controls in training cohort ( C ) and validation cohort ( D ). The “EJA” curve (blue line) shows healthy subjects vs. all EJA subjects (early + late-stage), and the “early-stage EJA” curve (green line) shows healthy subjects vs. only the early-stage EJA subjects. EJA esophagogastric junction adenocarcinoma, ROC receiver-operating characteristics curve

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques: Biomarker Discovery

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Diagnostic results of serum IGFBP3 in EJA

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques: Diagnostic Assay, Biomarker Discovery

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Kaplan–Meier curves for serum IGFBP3 with EJA patients. Patients with low serum IGFBP3 level had short survival. EJA esophagogastric junction adenocarcinoma

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques:

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Construction and performance evaluation of the nomogram. Nomogram based on serum IGFBP3, tumor size and TNM stage to predict the 1-, 3- and 5-years OS for EJA patients. The nomogram used by adding the points determined on the point scale of each variable, the total points projected on the bottom scales match the probability of 1-, 3-, and 5-year OS of EJA patient ( A ). Time-dependent C-index of nomogram based on serum IGFBP3, tumor size and TNM stage for OS of EJA patient ( B ) and internally validated with using a bootstrap resampling method ( C ). Calibration plots showed good agreement for the survival probability at 1-, 3- and 5-year between the nomogram prediction and actual observed results ( D – F ). Decision curve analysis of serum IGFBP3, tumor size and TNM stage and nomogram. The straight black line represents the assumption that all patients die, and the horizontal line represents the assumption that no deaths happen ( G ). Clinical impact curve of nomogram. The red curve (number of high-risk individuals) indicated the number of people who were classifed as positive (high risk) by the nomogram at each threshold probability; the blue dotted curve (number of high-risk individuals with event) was the number of true positives at each threshold probability ( H ). EJA esophagogastric junction adenocarcinoma, OS overall survival

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques:

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Predictive improvement of the nomogram

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques: Significance Assay

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: Cox proportional hazards regression analysis of OS in EJA

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques: Significance Assay

Journal: Discover. Oncology

Article Title: Serum insulin-like growth factor binding protein 3 as a promising diagnostic and prognostic biomarker in esophagogastric junction adenocarcinoma

doi: 10.1007/s12672-022-00591-1

Figure Lengend Snippet: The C-index, AIC and BIC of prognostic factors and nomogram for prediction OS

Article Snippet: The IGFBP3 concentration in the serum was measured with a IGFBP3 ELISA Kit (CUSABIO, Wuhan, China).

Techniques: Significance Assay