Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human LRIG3 Antibody

LRIG3 antibody in Human Cervical Cancer Tissue by Immunohistochemistry (IHC-P).

LRIG3 in Human Cervical Cancer Tissue.

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.

Detection of LRIG3 by Western Blot

Detection of LRIG3 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Western Blot

Detection of LRIG3 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Western Blot

Detection of LRIG3 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Western Blot

Detection of LRIG3 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/33718179), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Immunohistochemistry

Detection of LRIG3 by Immunohistochemistry

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 (https://pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Immunohistochemistry

Detection of LRIG3 by Immunohistochemistry

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 (https://pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Western Blot

Detection of LRIG3 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LRIG3 by Western Blot

Detection of LRIG3 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/36639372), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human LRIG3 Antibody

Application
Recommended Usage

Immunohistochemistry

3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human cervical cancer tissue

Western Blot

0.1 µg/mL
Sample: Recombinant Human LRIG3 (Catalog # 3495-LR)

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.


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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: LRIG3

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.

References

  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.

Long Name

Leucine-rich Repeats and Immunoglobulin-like Domains 3

Alternate Names

LIG-3

Entrez Gene IDs

121227 (Human); 320398 (Mouse)

Gene Symbol

LRIG3

UniProt

Additional LRIG3 Products

Product Documents for Human LRIG3 Antibody

Certificate of Analysis

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Product Specific Notices for Human LRIG3 Antibody

For research use only

Related Research Areas

Citations for Human LRIG3 Antibody

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