Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate - Chlorocebus aethiops (African Green Monkey)

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IGFBP-3 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human IGFBP‑3
Gly28-Lys291
Accession # CAA46087

Specificity

Detects human IGFBP‑3 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross reactivity with recombinant mouse IGFBP-3 is observed, and less than 1% cross-reactivity with recombinant human (rh) IGFBP‑1, rhIGFBP‑2, and rhIGFBP‑4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human IGFBP‑3 Antibody

IGFBP‑3 antibody in Human Colon by Immunohistochemistry (IHC-P).

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.

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

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

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

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

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

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

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

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

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

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

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.

Applications for Human IGFBP‑3 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon

Western Blot

0.1 µg/mL
Sample: Recombinant Human IGFBP‑3 (Catalog # 675-B3)

Neutralization

Measured by its ability to neutralize IGFBP‑3 inhibition of IGF‑II-dependent proliferation in the MCF‑7 human breast cancer cell line [Karey, K.P. et al. (1988) Cancer Research 48:4083]. The Neutralization Dose (ND50) is typically 0.75-3.0 µg/mL in the presence of 0.2 µg/mL Recombinant Human IGFBP‑3 and 14 ng/mL Recombinant Human IGF‑II.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IGFBP-3

The superfamily of insulin-like growth factor (IGF) binding proteins include the six high-affinity IGF binding proteins (IGFBP) and at least four additional low-affinity binding proteins referred to as IGFBP related proteins (IGFBP-rP). All IGFBP superfamily members are cysteine-rich proteins with conserved cysteine residues, which are clustered in the amino- and carboxy-terminal thirds of the molecule. IGFBPs modulate the biological activities of IGF proteins. Some IGFBPs may also have intrinsic bioactivity that is independent of their ability to bind IGF proteins. Post-translational modifications of IGFBPs, including glycosylation, phosphorylation and proteolysis, have been shown to modify the affinities of the binding proteins to IGF.

Human IGFBP-3 cDNA encodes a 291 amino acid (aa) residue precursor protein with a putative 27 aa residue signal peptide that is processed to generate the 264 aa residue mature protein with three potential N-linked and two potential O-linked glycosylation sites. Human IGFBP-3 is expressed in multiple tissues. The highest expression level is found in the non-paranchymal cells of the liver. Expression levels are also higher during extrauterine life and peak during puberty. Human IGFBP-3 is the major IGF binding protein in plasma where it exists in a ternary complex with IGF-I or IGF-II and the acid-labile subunit (ALS).

References

  1. Jones, J.I. and D.R. Clemmons (1995) Endocrine Rev. 16:3.
  2. Kelley, K.M. et al. (1996) Int. J. Biochem. Cell Biol. 28:619.
  3. Spagnoli, A. and R.G. Rosenfeld (1997) Curr. Op. Endocrinology and Diabetes 4:1.

Long Name

Insulin-like Growth Factor Binding Protein 3

Alternate Names

IGFBP3

Entrez Gene IDs

3486 (Human); 16009 (Mouse)

Gene Symbol

IGFBP3

UniProt

CAA46087

Additional IGFBP-3 Products

Product Documents for Human IGFBP‑3 Antibody

Certificate of Analysis

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Product Specific Notices for Human IGFBP‑3 Antibody

For research use only

Citations for Human IGFBP‑3 Antibody

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Protocols

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