LRIG3 Antibody [Unconjugated]

LRIG3 was detected in immersion fixed paraffin-embedded sections of human cervical cancer tissue using Goat Anti-Human LRIG3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3495) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

LRIG3 inhibited glioma angiogenesis by regulating VEGFA expression. (A) qRT-PCR analysis targeting angiogenic factors in glioma cells transduced with LRIG3 and control siRNAs. (B) Western blot analysis of VEGFA, LRIG3, and GAPDH. (C) Integrated density of the bands was normalized to GAPDH in LRIG3 knockdown or overexpression glioma cells. (D) Levels of VEGFA protein in the CM from glioma cells with LRIG3 knockdown or overexpression were detected by ELISA. Data are means ± SD of 3 independent replicates. *p < 0.05; **p < 0.01; ***p < 0.001. CM, conditioned medium. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.

Clinical relevance of LRIG3, p-AKT, and VEGFA expression in gliomas. (A) IHC staining targeting LRIG3, p-AKT, and VEGFA in two representative GBM specimens. Brown staining, positive immunoreactivity. (B) Relative levels of LRIG3, p-AKT, and VEGFA proteins in GBM specimens (with low and high LRIG3 expression levels in 28 GBM patients). (C) Expression analysis (left) and correlation (right) between LRIG3 and VEGFA expression in 10 freshly collected human glioma samples. Data are means ± SD of 3 independent replicates. ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.

Ectopic expression of LRIG3 reduces the pro-angiogenic activity of glioma cells in vitro.(A) Western blots showing levels of LRIG3 expression in the vector control and LRIG3-transduced glioma cells. GAPDH was used as an internal control. (B) Representative images (left) and quantification (right) of wound healing assays in HUVECs treated with CM derived from the vector control or LRIG3-transduced glioma cells. (C) Representative images (left) and quantification (right) of transwell migration assays of HUVECs treated with the indicated CM. (D) Representative images (left) and quantification (right) of HUVECs formed tube-like structures on Matrigel-coated plates with CM derived from the vector control or LRIG3-transduced glioma cells. (E) HUVEC viability was determined using the MTT assay. HUVECs were treated with CM derived from the indicated cells for the specific number of days. Data are means ± SD of 3 replicates. *p < 0.05; **p < 0.01; ***p < 0.001. CM, conditioned medium. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.

Downregulation of LRIG3 enhances the pro-angiogenic activity of glioma cells in vitro. (A) Western blot analysis of LRIG3 expression in the siRNA control and LRIG3-silenced glioma cells. GAPDH was used as an internal control. (B) Representative images (left) and quantification (right) of wound healing assays of HUVECs treated with CM derived from the siRNA control or LRIG3-silenced glioma cells. (C) Representative images (left) and quantification (right) of transwell migration assays in HUVECs treated with the indicated CM. (D) Representative images (left) and quantification (right) of HUVECs formed tube-like structures on Matrigel-coated plates with CM derived from the siRNA control cells or LRIG3-silenced glioma cells. (E) HUVEC viability was determined using the MTT assay. HUVECs were treated with CM derived from the indicated cells for the specific number of days. Data are presented as means ± SD of 3 independent replicates. *p < 0.05; **p < 0.01. CM, conditioned medium. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.

LRIG3 expression is negatively correlated with tumor-supportive TAMs in human GBM and informs good prognosis of GBM patients. A, b The stroma score (a) and immune score (b) of LRIG3-High-expression and LRIG3-low-expression patients in the TCGA GBM database. The stroma and immune scores were determined based on expression data (Yoshihara et al., 2013). Unpaired student’s t test. c GSEA analysis based on KEGG gene sets and TCGA GBM database. d GSEA analysis for various types of immune cells in LRIG3-High-expression and LRIG3-low-expression patients in TCGA GBM database. e Representative images of HE staining and the low- and high-expression levels of LRIG3, CD163, and IBA1 in human GBM tissue microarrays. Scale bar, 50 mm. f, g Correlation analysis between LRIG3 and CD163 expression in TMA of low grade (grade 2 and 3) glioma (n = 31) (f) and GBM (n = 40) (g). Pearson’s correlation test. h Kaplan–Meier survival curves of LRIG3-High-expression and LRIG3-Low-expression patients in TCGA GBM database. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.

LRIG3 expression is negatively correlated with tumor-supportive TAMs in human GBM and informs good prognosis of GBM patients. A, b The stroma score (a) and immune score (b) of LRIG3-High-expression and LRIG3-low-expression patients in the TCGA GBM database. The stroma and immune scores were determined based on expression data (Yoshihara et al., 2013). Unpaired student’s t test. c GSEA analysis based on KEGG gene sets and TCGA GBM database. d GSEA analysis for various types of immune cells in LRIG3-High-expression and LRIG3-low-expression patients in TCGA GBM database. e Representative images of HE staining and the low- and high-expression levels of LRIG3, CD163, and IBA1 in human GBM tissue microarrays. Scale bar, 50 mm. f, g Correlation analysis between LRIG3 and CD163 expression in TMA of low grade (grade 2 and 3) glioma (n = 31) (f) and GBM (n = 40) (g). Pearson’s correlation test. h Kaplan–Meier survival curves of LRIG3-High-expression and LRIG3-Low-expression patients in TCGA GBM database. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.

ADAM17 promotes the secretion of soluble LRIG3 by glioma cells. A Immunoblots for LRIG3 in the cell lysates of human glioma cell lines (U118MG, U87MG, FU) and macrophage-like cell line (THP-1). b Immunoblots for LRIG3 in the cell lysates of mouse glioma cell line (GL261) and macrophage-like cell line (RAW264.7 and BMDM). c ELISA standard curve showing the correlation relationship of LRIG3 concentrations and the log of the optical density 490. d ELISA and immunoblots for the expression of soluble LRIG3 protein of supernatant medium (up panel), ADAM17, and LRIG3 in the cell lysates (down panel) in negative control and si-ADAM17 U118MG and GL261 cells. n = 6 biological replicates for ELISA assay. One-way ANOVA with bonferroni correction. e, f ELISA for the expression of soluble LRIG3 protein of supernatant medium in U118MG (e) and GL261 (f) cells with stimulation of ADAM17 agonist PMA, ADAM17 inhibitor TAPI and ADAM17 siRNAs. n = 6 biological replicates. One-way ANOVA with bonferroni correction. g, h ELISA for the expression of soluble LRIG3 protein of supernatant medium in U118MG (g) and GL261 (h) cells in the absence or presence of increasing doses of ADAM17 agonist PMA. n = 3 biological replicates. One-way ANOVA. i, j ELISA for the expression of soluble LRIG3 protein of supernatant medium in U118MG (i) and GL261 (j) cells in the absence or presence of increasing doses of ADAM17 inhibitor TAPI. n = 3 biological replicates. One-way ANOVA. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.

ADAM17 promotes the secretion of soluble LRIG3 by glioma cells. A Immunoblots for LRIG3 in the cell lysates of human glioma cell lines (U118MG, U87MG, FU) and macrophage-like cell line (THP-1). b Immunoblots for LRIG3 in the cell lysates of mouse glioma cell line (GL261) and macrophage-like cell line (RAW264.7 and BMDM). c ELISA standard curve showing the correlation relationship of LRIG3 concentrations and the log of the optical density 490. d ELISA and immunoblots for the expression of soluble LRIG3 protein of supernatant medium (up panel), ADAM17, and LRIG3 in the cell lysates (down panel) in negative control and si-ADAM17 U118MG and GL261 cells. n = 6 biological replicates for ELISA assay. One-way ANOVA with bonferroni correction. e, f ELISA for the expression of soluble LRIG3 protein of supernatant medium in U118MG (e) and GL261 (f) cells with stimulation of ADAM17 agonist PMA, ADAM17 inhibitor TAPI and ADAM17 siRNAs. n = 6 biological replicates. One-way ANOVA with bonferroni correction. g, h ELISA for the expression of soluble LRIG3 protein of supernatant medium in U118MG (g) and GL261 (h) cells in the absence or presence of increasing doses of ADAM17 agonist PMA. n = 3 biological replicates. One-way ANOVA. i, j ELISA for the expression of soluble LRIG3 protein of supernatant medium in U118MG (i) and GL261 (j) cells in the absence or presence of increasing doses of ADAM17 inhibitor TAPI. n = 3 biological replicates. One-way ANOVA. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

LRIG3 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human LRIG3 isoform 1 (R&D Systems, Catalog # 3495-LR)
  Asp28-Thr807
  Accession # Q6UXM1
• Specificity: Detects human LRIG3 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant mouse LRIG1 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: LRIG3
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for LRIG3 Antibody [Unconjugated]

• FLJ90440
• KIAA3016
• leucine-rich repeats and immunoglobulin-like domains 3
• leucine-rich repeats and immunoglobulin-like domains protein 3
• LIG-3
• LIG3FLJ26573
• LIRG3
• LRIG3

Background

LRIG3 (leucine-rich repeats and Ig-like domains-3) is a 140 kDa type I transmembrane glycoprotein member of the mammalian LRIG glycoprotein family. This family contains three members who share 45 - 50% amino acid (aa) identity (1). All members contain at least fifteen LRRs, accompanied by two flanking cysteine-rich regions, and three C2-type Ig-like domains in their extracellular domains (ECD) (1). LRIG3 mRNA is widely expressed, with highest levels in stomach, skin, thyroid and small intestine (1). Human LRIG3 is synthesized as a 1120 amino acid (aa) precursor. It contains a 24 aa signal sequence, a 786 aa ECD, a 21 aa transmembrane sequence, and a 289 aa intracellular region. One splice variant exists that has a 19 aa substitution for the first 79 aa of the standard (or long) form. This substitution appears to encode an alternate signal sequence, resulting in a mature protein that lacks the first and part of the second LRR. LRIG1, a related family member, is known to bind the EGF family receptors ErbB1-4, via either its LRR or Ig-like domains. It also binds the ubiquitin ligase, c-Cbl, and promotes ubiquitination, internalization and destruction of these receptors (2, 3). It is not known whether LRIG3 performs similar functions. Within the cell, LRIG3 is expressed in the perinuclear region as well as on the cell surface. Perinuclear location of LRIG3 in grade III and IV astrocytic tumors has been associated with better patient survival (4). Human LRIG3 ECD shows 91%, 92%, 95% and 98% aa identity with mouse, rat, bovine and canine LRIG3 ECD, respectively.

1. Guo, D. et al. (2004) Genomics 84:157.
2. Gur, G. et al. (2004) EMBO J. 23:3270.
3. Laederich, M.B. (2004) J. Biol. Chem. 279:47050.
4. Guo, D. et al. (2006) Acta Neuropathol. (Berl.) 111:238.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LRIG3 Antibody (AF3495)(3)

Publications using AF3495

• Chenghao Peng, Hanmin Chen, Youwei Li, Hang Yang, Peizhong Qin, Baojun Ma et al. LRIG3 Suppresses Angiogenesis by Regulating the PI3K/AKT/VEGFA Signaling Pathway in Glioma Frontiers in Oncology 2/25/2021 [PMID: 33718179]
• Fangling Cheng, Po Zhang, Qungen Xiao, Youwei Li, Minhai Dong, Heping Wang et al. The Prognostic and Therapeutic Potential of LRIG3 and Soluble LRIG3 in Glioblastoma Frontiers in Oncology 6/6/2019 [PMID: 31245283]
• Rafidi , Hanine, Mercado , Francisc, Astudillo , Michael, Fry , William, Saldana , Matthew, Carraway , Kermit L, Sweeney , Colleen Leucine-rich repeat and immunoglobulin domain-containing protein-1 (Lrig1) negative regulatory action toward ErbB receptor tyrosine kinases is opposed by leucine-rich repeat and immunoglobulin domain-containing protein 3 (Lrig3). J Biol Chem, 2013-05-30;288(30):21593-605. 2013-05-30 [PMID: 23723069] (Western Blot, Human)

Bioinformatics

• Gene Symbol: LRIG3
• Uniprot:
  • Q6UXM1()