mouse specific sandwich elisa kit  (Elabscience Biotechnology)

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: Cordycepin mitigates 2% DSS-induced damage in NCM460 cells. ( A ) Effects of different concentrations of DSS on the viability of NCM460 cells. ( B ) Effects of different concentrations of cordycepin (COR) on the viability of NCM460 cells. NCM460 cells were treated with 2% DSS and / or 1 μmol/L (COR-1), 10 μmol/L (COR-10) COR for 24 h, the release levels of ( C ) LDH, ( D ) IL-1β, ( E ) IL-6, and ( F ) TNF-α in the supernatant were measured by using Elisa. ( G ) Representative images of PI staining and the percentage of PI-positive cells, scar bar = 100 μm. ( H ) The protein expression of ZO-1 in NCM460 cells was determined by using Western blotting assay. The results were representative of three independent experiments and expressed as mean ± SEM. Data were compared using two-tailed Student’s t tests. # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 vs. control group; *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 vs. the 2% DSS group.

Article Snippet: Serum CRP levels were measured using a mouse-specific sandwich ELISA kit (E-EL-M0053, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Expressing, Western Blot, Two Tailed Test, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: Cordycepin abates DSS-induced damage in NCM460 cells by activating adenosine receptor A 2A . ( A ) The cAMP levels in NCM460 cells under treatment with 2% DSS and/or 1 μmol/L and 10 μmol/L COR. ( B ) The cAMP levels in NCM460 cells under treatment with 2% DSS, 10 μmol/L COR, 5 μmol/L SCH58261 (SCH), and / or 5 μmol/L DPCPX for 24 h. The release levels of ( C ) LDH, ( D ) IL-1β, ( E ) IL-6, and ( F ) TNF-α were measured by using Elisa. ( G ) Representative images of PI staining and the percentage of PI-positive cells, scar bar = 100 μm. ( H – J ) The protein expression of ZO-1, A 2A AR and A 1 AR in NCM460 cells was determined by using Western blotting assay. The results were representative of three independent experiments and expressed as mean ± SEM. Data were compared using two-tailed Student’s t tests. vs. control group; # P < 0.05, vs. control group; *P < 0.05, **P < 0.01 and ***P < 0.001 vs. the 2% DSS group.

Article Snippet: Serum CRP levels were measured using a mouse-specific sandwich ELISA kit (E-EL-M0053, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Expressing, Western Blot, Two Tailed Test, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: SCH58261 blocks cordycepin’s amelioration of intestinal inflammation and gut barrier function in DSS-induced colitis mice. ( A – C ) Level of colonic IL-1β, IL-6, and TNF-α among the Control, DSS, DSS+COR-L and DSS+COR-H group were detected by using Elisa. ( D ) Representative images of AB-PAS staining of colonic tissue. The arrow indicates the goblet cells. ( E ) Number of goblet cells in colonic tissue. ( F ) Representative images of TUNEL staining in colonic tissue, scale bar = 50 μm. ( G ) Percentage of TUNEL-positive cells (%). ( H ) The protein expression of ZO-1 and Occludin in colons was determined by using Western blotting assay. ( I ) and ( J ) Serum level of D-lactate and diamine oxidase (DAO) were measured by using Elisa. ( K ) The serum cAMP levels among the control, DSS, DSS+COR-L, DSS+COR-H group and DSS+SCH+COR-H group. Data were presented as the means ± SEM of six-eight mice in each group and were compared using two-tailed Student’s t tests. # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 vs. control group; *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 vs. the DSS model (DSS) group.

Article Snippet: Serum CRP levels were measured using a mouse-specific sandwich ELISA kit (E-EL-M0053, Elabscience).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Staining, TUNEL Assay, Expressing, Western Blot, Two Tailed Test

Journal: Cells

Article Title: Anti-Inflammatory Effects of C1q/Tumor Necrosis Factor-Related Protein 3 (CTRP3) in Endothelial Cells

doi: 10.3390/cells10082146

Figure Lengend Snippet: Tissue expression of Ctrp3. Ctrp3 (C1q/TNF-related protein 3) mRNA levels in different murine tissues and organs were determined by Real-time PCR and CTRP3 serum level by ELISA. n = 4–6 mice.

Article Snippet: Supernatant from MyEnd cells and murine blood serum was analyzed for TNF-α, sVCAM-1, and sICAM-1 applying mouse-specific ELISA kits from R&D Systems (Minneapolis, MN, USA) and CTRP3 was quantified in murine blood serum applying a mouse-specific ELISA kit (LSBio, Seattle, WA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Anti-Inflammatory Effects of C1q/Tumor Necrosis Factor-Related Protein 3 (CTRP3) in Endothelial Cells

doi: 10.3390/cells10082146

Figure Lengend Snippet: Characterization of recombinant CTRP3. ( A ) Indicated quantities of recombinant CTRP3 (C1q/TNF-related protein 3) were run on SDS-PAGE (left) and 1000 ng CTRP3 for a longer gel run (right). Gels were subsequently stained with Coomassie. M = protein size marker. ( B ) THP-1 monocyte-like cells were stimulated with LPS (10 ng/mL) alone or pretreated with CTRP3 (10 µg/mL, for 30 min) and with CTRP3 alone for 18 h. CCL2 (C-C motif chemokine ligand 2) and IL-6 (interleukin-6) protein levels in the cell supernatant were analyzed by ELISA ** p < 0.01 vs. con = unstimulated control, # p < 0.05 vs. LPS, n = 5–6.

Article Snippet: Supernatant from MyEnd cells and murine blood serum was analyzed for TNF-α, sVCAM-1, and sICAM-1 applying mouse-specific ELISA kits from R&D Systems (Minneapolis, MN, USA) and CTRP3 was quantified in murine blood serum applying a mouse-specific ELISA kit (LSBio, Seattle, WA, USA).

Techniques: Recombinant, SDS Page, Staining, Marker, Enzyme-linked Immunosorbent Assay, Control

Journal: Cells

Article Title: Anti-Inflammatory Effects of C1q/Tumor Necrosis Factor-Related Protein 3 (CTRP3) in Endothelial Cells

doi: 10.3390/cells10082146

Figure Lengend Snippet: LPS-induced endothelial adhesion molecule expression is inhibited by CTRP3. ( A ) MyEnd (myocardial endothelial) cells were stimulated with LPS (lipopolysaccharides, 50 ng/mL) for the indicated period. Vcam-1 (vascular cell adhesion molecule-1) and Icam-1 (intercellular adhesion molecule-1) mRNA levels were analyzed by real-time PCR. * p < 0.05, ** p < 0.01 vs. con = unstimulated control, n = 5–6. ( B ) MyEnd cells were stimulated with LPS (50 ng/mL) alone or pretreated with CTRP3 (C1q/TNF-related protein 3, 10 µg/mL, pre-incubation for 30 min) and with CTRP3 alone for 3 h; Vcam-1 and Icam-1 mRNA levels were analyzed by real-time PCR. * p < 0.05, ** p < 0.01 vs. con = unstimulated control, # p < 0.05, ## p < 0.01 vs. LPS, n = 4–6. ( C ) MyEnd cells were stimulated with LPS (50 ng/mL) alone or pretreated with CTRP3 (10 µg/mL, for 30 min) and with CTRP3 alone for 3 h. The sVCAM-1 (soluble vascular cell adhesion molecule-1) and sICAM-1 (soluble intercellular adhesion molecule-1) protein levels in the cell supernatant were analyzed by ELISA. * p < 0.05 vs. con = unstimulated control, # p < 0.05 vs. LPS, n = 3–5.

Article Snippet: Supernatant from MyEnd cells and murine blood serum was analyzed for TNF-α, sVCAM-1, and sICAM-1 applying mouse-specific ELISA kits from R&D Systems (Minneapolis, MN, USA) and CTRP3 was quantified in murine blood serum applying a mouse-specific ELISA kit (LSBio, Seattle, WA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Anti-Inflammatory Effects of C1q/Tumor Necrosis Factor-Related Protein 3 (CTRP3) in Endothelial Cells

doi: 10.3390/cells10082146

Figure Lengend Snippet: LPS-induced systemic markers of endothelial inflammation are not affected by CTRP3 in vivo. Male C57BL/6J wild-type mice were treated with i.p. injection of recombinant CTRP3 (C1q/TNF-related protein 3, 10 µg/animal) 30 min prior to i.p. LPS-injection (lipopolysaccharides, 1 µg/animal) for 2 h. LPS and CTRP3 alone were used as controls. ( A ) TNF-α (tumor necrosis factor-α), ( B ) sICAM-1 (soluble intercellular adhesion molecule-1) and ( C ) sVCAM-1 (soluble vascular cell adhesion molecule-1) protein levels in blood serum were quantified by ELISA. * p < 0.05 con = unstimulated control, n = 3–9.

Article Snippet: Supernatant from MyEnd cells and murine blood serum was analyzed for TNF-α, sVCAM-1, and sICAM-1 applying mouse-specific ELISA kits from R&D Systems (Minneapolis, MN, USA) and CTRP3 was quantified in murine blood serum applying a mouse-specific ELISA kit (LSBio, Seattle, WA, USA).

Techniques: In Vivo, Injection, Recombinant, Enzyme-linked Immunosorbent Assay, Control