quantikine elisa kit  (R&D Systems)

Journal: Journal of Bone Oncology

Article Title: Doxorubicin enhances adipogenesis in an FGF2-dependent manner and induces a tumour-promoting secretory phenotype

doi: 10.1016/j.jbo.2026.100754

Figure Lengend Snippet: Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by ELISA in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.

Article Snippet: ELISA to measure protein concentration of FGF2 in conditioned supernatants was performed using the Quantikine ELISA kit (cat #DFB50, R&D Systems, Minneapolis, MN), according to manufacturer’s instructions.

Techniques: Expressing, Control, High Molecular Weight, Molecular Weight, Enzyme-linked Immunosorbent Assay, Transfection, Quantitative RT-PCR, Staining, Isolation, Gene Expression

Journal: Stem Cell Research & Therapy

Article Title: Extracellular vesicles derived from clonal mesenchymal stromal cells preconditioned by indirect hypoxia modulate immune responses in diabetic mice more effectively than directly preconditioned vesicles

doi: 10.1186/s13287-025-04568-z

Figure Lengend Snippet: Schematic of direct and indirect preconditioning of hBM-cMSCs: Illustration of preconditioning regimens applied to hBM-cMSCs, including direct (dFN, dPO, dLP, dH) and indirect (iFN, iPO, iLP, iH) preconditioned cMSCs. Conditioned media (cMSC-CCM) from these groups were concentrated and evaluated for their immunomodulatory and regenerative potential. The selected groups were used for further EV isolation and in vivo studies. Abbreviations: BM: bone marrow, CM: conditioned medium, cMSC: clonal mesenchymal stromal cell, dFN: direct IFN-γ, dH: direct hypoxia, dLP: direct LPS, dPO: direct poly(I: C), ELISA: enzyme-linked immunosorbent assay, IFN-γ: interferon-gamma, iFN: indirect IFN-γ preconditioning, iH: indirect hypoxia, iLP: indirect LPS, iPO: indirect poly(I: C), LPA: lymphocyte proliferation assay, LPS: lipopolysaccharide, µg: microgram, ml: milliliter, ng: nanogram, PBMC: peripheral blood mononuclear cell, poly(I: C): polyinosinic-polycytidylic acid, WB: Western blotting

Article Snippet: To measure cytokines in both direct and indirect cMSC-CCM, the levels of IL-10, IL-6, IL-1β, TGF-β, IFN-γ, TNF-α, VEGF, and IGF-1 were measured via DuoSet ® ELISA kits (DY217B, DY206, DY201, DY240, DY285B, DY210, DY293B, DY291, R&D Systems, USA), and the amounts of HGF and bFGF were measured via human Quantikine ® ELISA kits (DHG00 and DFB50, R&D Systems, USA) according to the manufacturer’s instructions.

Techniques: Isolation, In Vivo, Enzyme-linked Immunosorbent Assay, Lymphocyte Proliferation Assay, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Extracellular vesicles derived from clonal mesenchymal stromal cells preconditioned by indirect hypoxia modulate immune responses in diabetic mice more effectively than directly preconditioned vesicles

doi: 10.1186/s13287-025-04568-z

Figure Lengend Snippet: Direct preconditioning effects on cMSCs: ( A ) ELISA results showing cytokine and growth factor levels in cMSC-CCM from different preconditioning regimens. Notable increases in anti-inflammatory (TGF-β1, IL-10) and regenerative (VEGF, FGF2, HGF) factors were observed, particularly in dH-MSC-CCM. ( B-C ) Western blot analysis revealed significant upregulation of COX2, VEGF, and HGF in dH-MSC-CCM (Full-length blots are presented in Supplementary Material ). ( D ) LPA: PBMCs were stimulated with PHA. Each bar shows the suppression of cMSC proliferation by 1 × 10 5 CFSE-labeled PBMCs in the presence of different preconditioning regimens. The LPA results revealed no significant suppression of PBMC proliferation across the directly preconditioned groups. The data are shown as the means ± SDs ( n = 3, *: p value < 0.05 , **: p value < 0.01 , ***: p value < 0.005 , and ****: p value < 0.001 ). Abbreviations: Ang1: Angiopoietin 1, bFGF (FGF-2): Basic fibroblast growth factor, CCM: Concentrated conditioned medium, cMSC: clonal mesenchymal stromal cell, COX-1: Cyclooxygenase-1, COX-2: Cyclooxygenase-2, ELISA: Enzyme-linked immunosorbent assay, FN: Interferon gamma-treated group, HGF: Hepatocyte growth factor, H: Hypoxia-treated group, IFN-γ: Interferon-gamma, IGF1: Insulin growth factor 1, IL-1β: Interleukin 1 beta, IL-4: Interleukin 4, IL-6: Interleukin 6, IL-10: Interleukin 10, JNK: c-Jun N-terminal kinase, kDa: Kilodalton, LP: LPS-treated group, LPA: Lymphocyte proliferation assay, µg: Microgram, MSC: Mesenchymal stromal cell, NF-κB: Nuclear factor kappa B, NO: Nontreated group, pg.: Picogram, PHA: Phytohemagglutinin, PO: poly(I: C)-treated group, SD: Standard deviation, LPA: Lymphocyte proliferation assay, µg: Microgram, TNF-α: Tumor necrosis factor alpha

Article Snippet: To measure cytokines in both direct and indirect cMSC-CCM, the levels of IL-10, IL-6, IL-1β, TGF-β, IFN-γ, TNF-α, VEGF, and IGF-1 were measured via DuoSet ® ELISA kits (DY217B, DY206, DY201, DY240, DY285B, DY210, DY293B, DY291, R&D Systems, USA), and the amounts of HGF and bFGF were measured via human Quantikine ® ELISA kits (DHG00 and DFB50, R&D Systems, USA) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Labeling, Lymphocyte Proliferation Assay, Standard Deviation

Journal: Stem Cell Research & Therapy

Article Title: Extracellular vesicles derived from clonal mesenchymal stromal cells preconditioned by indirect hypoxia modulate immune responses in diabetic mice more effectively than directly preconditioned vesicles

doi: 10.1186/s13287-025-04568-z

Figure Lengend Snippet: Indirect preconditioning effects on cMSCs: (A) ELISA results indicating greater levels of anti-inflammatory and regenerative cytokines in the iLP-MSC-CCM and iH-MSC-CCM groups than in the control group. ( B-C ) Western blot analysis revealed increased expression of COX2, VEGF, Ang1, and HGF in the iH-MSC-CCM group (Full-length blots are presented in Supplementary Material ). ( D ) LPA: PBMCs were stimulated with PHA. Each bar shows the suppression of cMSC proliferation by 1 × 10 5 CFSE-labeled PBMCs in the presence of different preconditioning regimens. LPA revealed that iLP-MSC-CCM and iH-MSC-CCM significantly inhibited PBMC proliferation, demonstrating strong immunomodulatory potential. The data are shown as the means ± SDs ( n = 3, *: p value < 0.05 , **: p value < 0.01 , ***: p value < 0.005 , and ****: p value < 0.001 ). Compared with the control, iLP-MSC-CCM and iH-MSC-CCM preconditioning resulted in almost 90% and 60% inhibition, respectively ( p value < 0.0001 )

Article Snippet: To measure cytokines in both direct and indirect cMSC-CCM, the levels of IL-10, IL-6, IL-1β, TGF-β, IFN-γ, TNF-α, VEGF, and IGF-1 were measured via DuoSet ® ELISA kits (DY217B, DY206, DY201, DY240, DY285B, DY210, DY293B, DY291, R&D Systems, USA), and the amounts of HGF and bFGF were measured via human Quantikine ® ELISA kits (DHG00 and DFB50, R&D Systems, USA) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing, Labeling, Inhibition

Journal: Stem Cell Research & Therapy

Article Title: Extracellular vesicles derived from clonal mesenchymal stromal cells preconditioned by indirect hypoxia modulate immune responses in diabetic mice more effectively than directly preconditioned vesicles

doi: 10.1186/s13287-025-04568-z

Figure Lengend Snippet: Effects of Hypoxia-Induced cMSC-EVs on MLD/STZ-Induced T1D Mice: ( A ) Timeline of the study outlining the induction of diabetes in mice and subsequent treatments with dH-MSCs, iH-MSCs, dH-MSC-EVs, and iH-MSC-EVs. ( B ) FBG levels were significantly lower in all treatment groups than in the sham group. ( C ) Kaplan‒Meier survival analysis revealed improved survival rates, particularly in the iH-MSC-EV group. ( D-G ) Analysis of serum cytokines revealed increased anti-inflammatory cytokine (IL-10) and decreased proinflammatory cytokine (IL-6, TNF-α) levels in iH-MSC-EV-treated mice. All the data are presented as the means ± SDs. ( *: p value < 0.05; **: p value < 0.01; ***: p value < 0.005; ****: p value < 0.001 ). Abbreviations: ANOVA: analysis of variance, cMSC: clonal mesenchymal stromal cell, dl: deciliter, DPBS: Dulbecco’s phosphate-buffered saline, ELISA: enzyme-linked immunosorbent assay, EV: extracellular vesicle, FBG: fasting blood glucose, gr: gram, h: hour, HbA1c: hemoglobin A1c, HSA: human serum albumin, H: hypoxia-treated group, IL-4: interleukin 4, IL-6: interleukin 6, IL-10: interleukin 10, IP: intraperitoneal injection, EV-GTT: intraperitoneal glucose tolerance test, IV: intravenous injection, kg: kilogram, µg: microgram, mg: milligram, MLD: multiple low-dose, µg: microgram, µl: microliter, MSC: mesenchymal stromal cell, N.S.: normal saline, PBMC/PB: peripheral blood mononuclear cell, pg: picogram, SD: standard deviation, STZ: Streptozotocin, TNF-α: tumor necrosis factor-alpha, Tx: treatment

Article Snippet: To measure cytokines in both direct and indirect cMSC-CCM, the levels of IL-10, IL-6, IL-1β, TGF-β, IFN-γ, TNF-α, VEGF, and IGF-1 were measured via DuoSet ® ELISA kits (DY217B, DY206, DY201, DY240, DY285B, DY210, DY293B, DY291, R&D Systems, USA), and the amounts of HGF and bFGF were measured via human Quantikine ® ELISA kits (DHG00 and DFB50, R&D Systems, USA) according to the manufacturer’s instructions.

Techniques: Saline, Enzyme-linked Immunosorbent Assay, Injection, Standard Deviation