anti human ttp  (Santa Cruz Biotechnology)

Journal: BioMed Research International

Article Title: Ultrashort Wave Combined with Human Umbilical Cord Mesenchymal Stem Cell (HUC-MSC) Transplantation Inhibits NLRP3 Inflammasome and Improves Spinal Cord Injury via MK2/TTP Signalling Pathway

doi: 10.1155/2020/3021750

Figure Lengend Snippet: MK2 upregulation blocks the microglia polarisation promoted by combination therapy: (a) iNOS and arg-1 expression levels detected using Western Blotting; (b) contents of inflammatory factors (IL-6 and IL-10) determined using ELISA; (c) Western Blotting was performed on to determine the expression levels of TTP, NLRP3, pro-IL-1 β , and pro-IL-18; (d) contents of IL-1 β and IL-18 in cellular supernatant were quantified with ELISA. “ ∗ , #, &” indicated significant difference ( p < 0.05), “ ∗ ” vs. Sham group, “#” vs. SCI group, and “&” vs. rats treated with monotherapies and combination therapy.

Article Snippet: Membranes were then incubated with diluted primary antibodies against MK2 (ab131531, Abcam, UK), p-MK2 (ab131504, Abcam, UK), TTP (ab124024, Abcam, UK), NLRP3 (ab263899, Abcam, UK), pro-caspase-1 (ab179515, Abcam, UK), pro-IL-1 β (ab205924, Abcam, UK), iNOS (ab15323, Abcam, UK), arg-1 (ab272887, Abcam, UK), and GAPDH (ab8245, Abcam, UK) as loading internal control at 4°C for 12 h. On the next day, membranes were incubated with HRP-conjugated secondary antibody for 1 h at room temperature after rinse with TBST buffer solution for 3 changes.

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

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Expression of human α-tocopherol transfer protein (α-TTP) eukaryotic expression vector in Human Hepatoma cells (HepG2). ( A ) Result of quantitative real-time PCR; ( B ) Result of Western blot; ( C ) Result of Immunofluorescence (10×). pcDNA: pcDNA3.1-mycHisa empty vector; p-TTP: human α-TTP eukaryotic expression vector.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Expression levels of α-TTP related genes. SEC23A : Sec23 homolog A; RTP4 : receptor transporter protein 4; CLIC3 : chloride intracellular channel 3; CENPE : centromere protein E; KCNQ1 : potassium voltage-gated channel subfamily Q member 1; GOLGA4 : golgi autoantigen, golgin subfamily a, 4; BNIP3 : BCL2/adenovirus E1B 19 kDa interacting protein 3; CENPF : centromere protein F. Data are presented as mean ± standard error; the asterisk indicated significant difference between control and treatment group.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Composition of the ligation reaction.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Ligation, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2)

doi: 10.3390/ijms17071016

Figure Lengend Snippet: Details of primers used for quantitative real-time PCR.

Article Snippet: The primary antibody was human α-TTP monoclonal (Abnova, Taiwan, Dilution ratio 1:1000) and the secondary antibody was goat anti-mouse (Dilution ratio 1:5000) (from Beijing B & M Biotech Co., Ltd., Beijing, China).

Techniques: Sequencing

Journal: Molecular Genetics and Metabolism Reports

Article Title: Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells

doi: 10.1016/j.ymgmr.2016.02.004

Figure Lengend Snippet: TTP overexpression represses ERα, PR, GR and AR transactivation in MCF-7 cells. Human breast cancer MCF-7 cells were transiently transfected with empty pCMV-3TAG vector (control) or with 0.25 μg of pCMV-3TAG-TTP (FLAG-TTP) along with the corresponding luciferase reporter vector. The effect of TTP on nuclear hormone receptors transactivation was determined by assay of luciferase activity, as described under “Experimental Procedures.” Assays were performed in triplicate in three independent experiments in the presence (white bars) or absence (black bars) of the corresponding nuclear receptor ligands (17β-estradiol for ERα, progesterone for PR, 5α-Androstan-17β-ol-3-on for AR and dexamethasone for GR. The results are represented as mean ± S.E. Differences in ERα activity between control MCF-7 cells and TTP-expressing MCF-7 cells are statistically significant ( p < 0.05).

Article Snippet: Human ERα antibody (Sc D -12, Sc HC-20) and Human TTP antibody (Sc-12565) were purchased from Santa Cruz Biotechnology, Inc. and TTP polyclonal (T5327) antibody was from Sigma-Aldrich.

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Luciferase, Activity Assay, Expressing

Journal: Molecular Genetics and Metabolism Reports

Article Title: Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells

doi: 10.1016/j.ymgmr.2016.02.004

Figure Lengend Snippet: TTP interacts with ERα, PR, GR and AR in vivo . Endogenous ERα (A), PR (B), AR (C) and GR (C) were immunoprecipitated from protein extracts prepared from MCF-7 cells using specific antibodies. Immunoprecipitated proteins were resolved by PAGE, and the binding of TTP was visualized by Western blot. The input lane represents 10% of the protein extract used in the coimmunoprecipitation assays. IP: immunoprecipitation, WB: Western blot.

Article Snippet: Human ERα antibody (Sc D -12, Sc HC-20) and Human TTP antibody (Sc-12565) were purchased from Santa Cruz Biotechnology, Inc. and TTP polyclonal (T5327) antibody was from Sigma-Aldrich.

Techniques: In Vivo, Immunoprecipitation, Binding Assay, Western Blot

Journal: Molecular Genetics and Metabolism Reports

Article Title: Nuclear tristetraprolin acts as a corepressor of multiple steroid nuclear receptors in breast cancer cells

doi: 10.1016/j.ymgmr.2016.02.004

Figure Lengend Snippet: siRNA-mediated knockdown of TTP protein levels in MCF-7 cells. MCF-7 cells were transfected with specific siRNA-TTP (1.25 μg) or with an unrelated control siRNA (2.5 μg). (A) Protein extracts from MCF-7 cultures were resolved by PAGE, and expression levels of TTP, ERα, PR, GR, AR and tubulin, as a loading control protein, were evaluated by Western blot using specific antibodies as described under “Experimental Procedures.” TTP knockdown increases ERα, PR, GR and AR transactivation. MCF-7 cells were transfected with an unrelated siRNA (control) or a specific TTP-siRNA (1.5 μg), along with TK-LUC, in the presence of the corresponding nuclear receptor ligands. The effect of TTP-siRNA on ERα (B), PR (C), GR (D) and AR (E) transactivation was determined by a luciferase assay and compared with luciferase activity in MCF-7 cells transfected with empty pCMV-3TAG vector and the corresponding Tk-LUC reporter vector. Results, in triplicate in three independent experiments, are represented as mean ± S.E. (error bars). Differences in ERα activity in MCF-7 cells transfected with TTP or with TTP-siRNA were shown to be statistically significant ( p < 0.05).

Article Snippet: Human ERα antibody (Sc D -12, Sc HC-20) and Human TTP antibody (Sc-12565) were purchased from Santa Cruz Biotechnology, Inc. and TTP polyclonal (T5327) antibody was from Sigma-Aldrich.

Techniques: Knockdown, Transfection, Control, Expressing, Western Blot, Luciferase, Activity Assay, Plasmid Preparation