Human IGFBP-3 Antibody

IGFBP‑3 in Human Colon. IGFBP‑3 was detected in immersion fixed paraffin-embedded sections of human colon using 10 µg/mL Human IGFBP‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF675) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

IGFBP‑3 Inhibition of IGF‑II-dependent Cell Proliferation and Neutralization by Human IGFBP‑3 Antibody. Recombinant Human IGFBP‑3 (Catalog # 675-B3) inhibits Recombinant Human IGF‑II (Catalog # 292-G2) induced proliferation in the MCF‑7 human breast cancer cell line in a dose-dependent manner (orange line). Inhibition of Recombinant Human IGF‑II (14 ng/mL) activity elicited by Recombinant Human IGFBP‑3 (0.2 µg/mL) is neutralized (green line) by increasing concentrations of Human IGFBP‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF675). The ND₅₀ is typically 0.75-3.0 µg/mL.

Detection of IGFBP-3 by Western Blot IGFBP3 increases PD-L1 expression by up-regulating the phosphorylation of STAT3 in GBM cells. A KEGG analysis of differential genes. B Correlation analysis of IGFBP3 and STAT3 in TCGA and CGGA databases, and statistical significance was assessed using the spearman’s rank test. C Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cell treated with different concentrations of IGFBP3 for 24 h. D Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cells infected with IGFBP3 overexpressing and empty vector lentivirus. E Immunoblotting was performed to examine the expression of IGFBP3, p-JAK2, p-STAT3, and PD-L1 in LN229 cell transfected with si-IGFBP3 and si-NC. F Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in LN229 cell infected with sh-IGFBP3 and sh-con. G LN229 and T98G cells with IGFBP3 knockdown were treated with the JAK2/STAT3 inhibitor WP1066 for 24 h, cell lysate was used to analyze the expression of p-JAK2, p-STAT3 and PD-L1. H Immunoblotting was used to analyze the expression of IGFBP3, p-STAT3, and PD-L1 in STAT3-silenced LN229 and T98G cells stimulated with different concentrations of exogenous IGFBP3 for 24 h Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 transcriptionally regulates PD-L1 expression in GBM cells. A Immunoblotting analysis of IGFBP3 and PD-L1 expression in U251 and U118 cells treated with different concentrations of exogenous IGFBP3 (0, 25, 50, and 100 ng/mL) for 24 h. B Immunoblotting was performed to determine the expression of IGFBP3 and PD-L1 in U251 and U118 cells stimulated with 100 ng/mL of IGFBP3 for 0, 12, 24, and 48 h. C QRT-PCR analysis of IGFBP3 and PD-L1 mRNA expression levels in U251 and U118 cells infected with overexpressing IGFBP3 and empty vector lentivirus. D Immunoblotting assay of IGFBP3 and PD-L1 protein expression in U251 and U118 cells infected with overexpressing IGFBP3 and empty vector lentivirus. E Representative immunofluorescence images of DAPI (blue), IGFBP3 (red), PD-L1 (green) in U251-vector and U251-IGFBP3 cells. F Immunoblotting was performed to examine the expression of IGFBP3 and PD-L1 in LN229 and T98G cells transfected with si-RNA targeting IGFBP3 (si-IGFBP3) and negative control (si-NC). G Immunoblotting analysis of IGFBP3 and PD-L1 expression in LN229 and T98G cells infected with lentivirus of targeting IGFBP3 (sh-IGFBP3) and control (sh-con). Data are expressed as mean ± SD. **P < 0.01; ***P < 0.001, Student t test Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Immunocytochemistry/ Immunofluorescence Targeting IGFBP3 therapy efficiently suppresses glioblastoma invasion in vitro and tumor growth in vivo via reducing PD-L1 expression. A Immunoblotting analysis of IGFBP3 expression in GL261 cell infected with sh-IGFBP3 and sh-con. B Immunofluorescence was used to examine IGFBP3 expression in GL261 cell infected with sh-IGFBP3 and sh-con. C Left: Representative image of brain orthotopic tumor. Right: HE stained representative image of brain tumor section. D Kaplan–Meier plots for GL261-shcon and GL261-shIGFBP3 groups, and statistical significance was assessed using the log-rank test. E Immunohistochemical representative images of IGFBP3, PD-L1 and CD31 in the GL261-shcon and GL261-shIGFBP3 groups. F IGFBP3 up-regulates the expression of PD-L1 by activating JAK2/STAT3 signaling pathway. PD-L1 binds to T cell–derived PD-1 to promote T cell apoptosis and immunosuppression. Data are expressed as mean ± SD. **P < 0.01; ***P < 0.001, Student t test Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 increases PD-L1 expression by up-regulating the phosphorylation of STAT3 in GBM cells. A KEGG analysis of differential genes. B Correlation analysis of IGFBP3 and STAT3 in TCGA and CGGA databases, and statistical significance was assessed using the spearman’s rank test. C Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cell treated with different concentrations of IGFBP3 for 24 h. D Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cells infected with IGFBP3 overexpressing and empty vector lentivirus. E Immunoblotting was performed to examine the expression of IGFBP3, p-JAK2, p-STAT3, and PD-L1 in LN229 cell transfected with si-IGFBP3 and si-NC. F Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in LN229 cell infected with sh-IGFBP3 and sh-con. G LN229 and T98G cells with IGFBP3 knockdown were treated with the JAK2/STAT3 inhibitor WP1066 for 24 h, cell lysate was used to analyze the expression of p-JAK2, p-STAT3 and PD-L1. H Immunoblotting was used to analyze the expression of IGFBP3, p-STAT3, and PD-L1 in STAT3-silenced LN229 and T98G cells stimulated with different concentrations of exogenous IGFBP3 for 24 h Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 induces immunosuppression in in-vitro co-culture system. A CCK-8 assay was used to analysis the viability of Jurkat cells after co-cultured with IGFBP3 overexpressing cells. B, C QRT-PCR was performed to examine the mRNA expression of IL2 (B) or IFN-gamma (C) in Jurkat cells after co-cultured with IGFBP3 overexpressing cells. D, E ELISA was used to measure the levels of IL-2 (D) and IFN-gamma (E) in the supernatants of Jurkat cells after co-cultured with IGFBP3 overexpressing cells. F Annexin V-PE apoptosis assay was used to determine apoptosis of Jurkat cells after co-cultured with IGFBP3 overexpressing cells. G Statistical chart of apoptotic cells. H Immunoblotting was performed to examine the caspase3 expression in Jurkat cells after co-cultured with IGFBP3 overexpressing cells. Data are expressed as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001, Student t test Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 transcriptionally regulates PD-L1 expression in GBM cells. A Immunoblotting analysis of IGFBP3 and PD-L1 expression in U251 and U118 cells treated with different concentrations of exogenous IGFBP3 (0, 25, 50, and 100 ng/mL) for 24 h. B Immunoblotting was performed to determine the expression of IGFBP3 and PD-L1 in U251 and U118 cells stimulated with 100 ng/mL of IGFBP3 for 0, 12, 24, and 48 h. C QRT-PCR analysis of IGFBP3 and PD-L1 mRNA expression levels in U251 and U118 cells infected with overexpressing IGFBP3 and empty vector lentivirus. D Immunoblotting assay of IGFBP3 and PD-L1 protein expression in U251 and U118 cells infected with overexpressing IGFBP3 and empty vector lentivirus. E Representative immunofluorescence images of DAPI (blue), IGFBP3 (red), PD-L1 (green) in U251-vector and U251-IGFBP3 cells. F Immunoblotting was performed to examine the expression of IGFBP3 and PD-L1 in LN229 and T98G cells transfected with si-RNA targeting IGFBP3 (si-IGFBP3) and negative control (si-NC). G Immunoblotting analysis of IGFBP3 and PD-L1 expression in LN229 and T98G cells infected with lentivirus of targeting IGFBP3 (sh-IGFBP3) and control (sh-con). Data are expressed as mean ± SD. **P < 0.01; ***P < 0.001, Student t test Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 increases PD-L1 expression by up-regulating the phosphorylation of STAT3 in GBM cells. A KEGG analysis of differential genes. B Correlation analysis of IGFBP3 and STAT3 in TCGA and CGGA databases, and statistical significance was assessed using the spearman’s rank test. C Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cell treated with different concentrations of IGFBP3 for 24 h. D Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cells infected with IGFBP3 overexpressing and empty vector lentivirus. E Immunoblotting was performed to examine the expression of IGFBP3, p-JAK2, p-STAT3, and PD-L1 in LN229 cell transfected with si-IGFBP3 and si-NC. F Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in LN229 cell infected with sh-IGFBP3 and sh-con. G LN229 and T98G cells with IGFBP3 knockdown were treated with the JAK2/STAT3 inhibitor WP1066 for 24 h, cell lysate was used to analyze the expression of p-JAK2, p-STAT3 and PD-L1. H Immunoblotting was used to analyze the expression of IGFBP3, p-STAT3, and PD-L1 in STAT3-silenced LN229 and T98G cells stimulated with different concentrations of exogenous IGFBP3 for 24 h Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot Targeting IGFBP3 therapy efficiently suppresses glioblastoma invasion in vitro and tumor growth in vivo via reducing PD-L1 expression. A Immunoblotting analysis of IGFBP3 expression in GL261 cell infected with sh-IGFBP3 and sh-con. B Immunofluorescence was used to examine IGFBP3 expression in GL261 cell infected with sh-IGFBP3 and sh-con. C Left: Representative image of brain orthotopic tumor. Right: HE stained representative image of brain tumor section. D Kaplan–Meier plots for GL261-shcon and GL261-shIGFBP3 groups, and statistical significance was assessed using the log-rank test. E Immunohistochemical representative images of IGFBP3, PD-L1 and CD31 in the GL261-shcon and GL261-shIGFBP3 groups. F IGFBP3 up-regulates the expression of PD-L1 by activating JAK2/STAT3 signaling pathway. PD-L1 binds to T cell–derived PD-1 to promote T cell apoptosis and immunosuppression. Data are expressed as mean ± SD. **P < 0.01; ***P < 0.001, Student t test Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 transcriptionally regulates PD-L1 expression in GBM cells. A Immunoblotting analysis of IGFBP3 and PD-L1 expression in U251 and U118 cells treated with different concentrations of exogenous IGFBP3 (0, 25, 50, and 100 ng/mL) for 24 h. B Immunoblotting was performed to determine the expression of IGFBP3 and PD-L1 in U251 and U118 cells stimulated with 100 ng/mL of IGFBP3 for 0, 12, 24, and 48 h. C QRT-PCR analysis of IGFBP3 and PD-L1 mRNA expression levels in U251 and U118 cells infected with overexpressing IGFBP3 and empty vector lentivirus. D Immunoblotting assay of IGFBP3 and PD-L1 protein expression in U251 and U118 cells infected with overexpressing IGFBP3 and empty vector lentivirus. E Representative immunofluorescence images of DAPI (blue), IGFBP3 (red), PD-L1 (green) in U251-vector and U251-IGFBP3 cells. F Immunoblotting was performed to examine the expression of IGFBP3 and PD-L1 in LN229 and T98G cells transfected with si-RNA targeting IGFBP3 (si-IGFBP3) and negative control (si-NC). G Immunoblotting analysis of IGFBP3 and PD-L1 expression in LN229 and T98G cells infected with lentivirus of targeting IGFBP3 (sh-IGFBP3) and control (sh-con). Data are expressed as mean ± SD. **P < 0.01; ***P < 0.001, Student t test Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IGFBP-3 by Western Blot IGFBP3 increases PD-L1 expression by up-regulating the phosphorylation of STAT3 in GBM cells. A KEGG analysis of differential genes. B Correlation analysis of IGFBP3 and STAT3 in TCGA and CGGA databases, and statistical significance was assessed using the spearman’s rank test. C Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cell treated with different concentrations of IGFBP3 for 24 h. D Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in U251 cells infected with IGFBP3 overexpressing and empty vector lentivirus. E Immunoblotting was performed to examine the expression of IGFBP3, p-JAK2, p-STAT3, and PD-L1 in LN229 cell transfected with si-IGFBP3 and si-NC. F Immunoblotting analysis of IGFBP3, p-JAK2, p-STAT3, and PD-L1 expression in LN229 cell infected with sh-IGFBP3 and sh-con. G LN229 and T98G cells with IGFBP3 knockdown were treated with the JAK2/STAT3 inhibitor WP1066 for 24 h, cell lysate was used to analyze the expression of p-JAK2, p-STAT3 and PD-L1. H Immunoblotting was used to analyze the expression of IGFBP3, p-STAT3, and PD-L1 in STAT3-silenced LN229 and T98G cells stimulated with different concentrations of exogenous IGFBP3 for 24 h Image collected and cropped by CiteAb from the following open publication (https://cancerci.biomedcentral.com/articles/10.1186/s12935-024-03234-3), licensed under a CC-BY license. Not internally tested by R&D Systems.