Mouse Lgr5/GPR49 Antibody (803420)

R&D Systems | Catalog # MAB8240

Published as Clone RD20
R&D Systems
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Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse, Transgenic Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry, Agonist Activity, CyTOF-ready

Cited:

Immunohistochemistry, Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2B Clone # 803420
View all Lgr5/GPR49 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse Lgr5/GPR49
Lys375-Pro547
Accession # Q9Z1P4

Specificity

Detects mouse Lgr5/GPR49 in ELISA. Stains mouse Lgr5 transfected cells but not irrelevant transfectants in Flow Cytometry and Immunocytochemistry. This antibody, also known as "RD20", has been found to detect an epitope in C-terminal LRR cap of LGR5 (Ref. 1).

Clonality

Monoclonal

Host

Rat

Isotype

IgG2B

Scientific Data Images for Mouse Lgr5/GPR49 Antibody (803420)

Detection of Lgr5/GPR49 antibody in HEK293 Human Cell Line Transfected with Mouse Lgr5/GPR49 and eGFP antibody by Flow Cytometry.

Detection of Lgr5/GPR49 in HEK293 Human Cell Line Transfected with Mouse Lgr5/GPR49 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with mouse Lgr5/GPR49 and eGFP was stained with and either (A) Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240) or (B) Rat IgG2BIsotype Control (Catalog # MAB0061) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113).
Lgr5/GPR49 antibody in HEK293 Human Cell Line Transfected with Mouse Lgf5/GPR49 by Immunocytochemistry (ICC).

Lgr5/GPR49 in HEK293 Human Cell Line Transfected with Mouse Lgf5/GPR49.

Lgr5/GPR49 was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with GFP (green) tagged mouse LGR5 using Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Lgr5/GPR49 antibody in Mouse Cells Transfected with Mouse Lgr5/GPR49 by Immunocytochemistry (ICC).

Lgr5/GPR49 in Mouse Cells Transfected with Mouse Lgr5/GPR49.

Lgr5/GPR49 was detected in immersion fixed MYC tagged mouse transfectants using Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240). Cells were stained using an anti-Mouse HRP-conjugated secondary antibody (brown).Image courtesy of Dr. Hans Clevers, Hubrecht Institute, The Netherlands.
Mouse Lgr5/GPR49 Antibody Induces Activity.

Mouse Lgr5/GPR49 Antibody Induces Activity.

Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240) induces TOPflash activity in the HEK293 human embryonic kidney cell line stably expressing LGR5 in the presence of Wnt-3a, but in the absence of R-Spondins. (Data courtesy of Dr. Wim de Lau and Dr. Hans Clevers, Hubrecht Institute, The Netherlands. See Reference 1.)
Detection of fjhazglu7mkt2a2vqjxe.jpg by Immunohistochemistry

Detection of fjhazglu7mkt2a2vqjxe.jpg by Immunohistochemistry

LGR5 is a suitable marker for detecting stem cells in the rectal crypts. Immunofluorescence staining of the rectum and the jejunum of ICR wild-type mice for (A) co-staining of LGR5 (green) and ASCL2 (red) and (B) co-staining of LGR5 (green) and OLFM4 (red). The upper panel is for the rectum (test tissue) and the lower is for the jejunum (positive control). Positive signal marked in white boxes. Magnification 20×. Scale bar: 100 μm. Achaete–scute complex homolog 2 (ASCL2); leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5); olfactomedin 4 (OLFM4). Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/25/20/11252), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Lgr5/GPR49 by Immunohistochemistry

Detection of Lgr5/GPR49 by Immunohistochemistry

TT-2 promoted intestinal stem cell proliferation of mice after 14 Gy ABI. (A) Representative images of in situ hybridization for Lgr5 mRNA in intestinal crypts after 14 Gy ABI and different treatments (**p < 0.01, scale bar = 100 μm). Three mice from each group were used. (B) Representative Sox9-stained sections and quantification of Sox9+ cells in crypt of small intestine after 14 Gy ABI and different treatments (**p < 0.01, scale bar = 100 μm). Three mice from each group were used. (C) qPCR for Lgr5, Sox9, Dclk1, Hopx, Tert, and Prox1 gene expression in mice after 14 Gy irradiation with or without TT-2 treatment for 96 h after ABI (**p < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34796175), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Lgr5/GPR49 by Immunohistochemistry

Detection of Lgr5/GPR49 by Immunohistochemistry

ICR wild-type mice show faster recovery compared to Lgr5-EGFP knock-in mice. (A) Representative images of reappearances of the LGR5 population post-irradiation (10 days to 60 days). Arrows indicate representative clustered populations. Dashed squares indicate representative dotted populations. Days versus LGR5-positive stem cell relation in (B) 20 Gy CHBI, (C) 29 Gy CHBI. (D) Crypt microcolony survival assay. (E) Determining threshold dose after 30 days. Data represent the mean ± standard deviation. Asterisks indicate a significant difference (* p < 0.05, ** p < 0.01) between 0 days and the subsequent time points (9 days, 20 days, 30 days, 45 days and 60 days) and between the unirradiated group and other irradiated groups. Magnification 20×. Scale bar: 100 μm. Caudal half-body irradiation (CHBI); enhanced green fluorescent protein (EGFP); gray (Gy); leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5). Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/25/20/11252), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Lgr5/GPR49 by Immunohistochemistry

Detection of Lgr5/GPR49 by Immunohistochemistry

LGR5 is a suitable marker for detecting stem cells in the rectal crypts. Immunofluorescence staining of the rectum and the jejunum of ICR wild-type mice for (A) co-staining of LGR5 (green) and ASCL2 (red) and (B) co-staining of LGR5 (green) and OLFM4 (red). The upper panel is for the rectum (test tissue) and the lower is for the jejunum (positive control). Positive signal marked in white boxes. Magnification 20×. Scale bar: 100 μm. Achaete–scute complex homolog 2 (ASCL2); leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5); olfactomedin 4 (OLFM4). Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/25/20/11252), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse Lgr5/GPR49 Antibody (803420)

Application
Recommended Usage

Agonist Activity

Peng, W.C., et al. (2013) Cell Reports. 3:1885.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: HEK293 human embryonic kidney cell line transfected with mouse Lgr5/GPR49 and eGFP

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cell line transfected with GFP tagged mouse LGR5

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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: Lgr5/GPR49

Leucine‑rich repeat G‑protein‑coupled receptor 5 (Lgr5), also called GPR49, is a 907 amino acid (aa), approximately 97 kDa (calculated), seven‑transmembrane glycoprotein receptor in the Lgr family of cell surface receptors. The subfamily of Lgrs comprising Lgr4, Lgr5, and Lgr6 are G‑protein‑independent mediators of the potentiating effect of R‑Spondins on Wnt signaling (2). Lgr5 binds and forms complexes with R‑Spondins, Frizzled Wnt receptors and LRP Wnt co‑receptors. The region of the mouse Lgr5 long extracellular domain used as an immunogen shares 90% and 95% amino acid sequence identity with human and rat Lgr5, respectively. Lgr5 is found on embryonic and adult epithelial stem cells (3). Lgr5+ stem cells can produce all epithelial cell types of the intestinal crypts (4). It is upregulated in stem cells that give rise to cancers such as intestinal, hepatocellular, pancreatic and ovarian carcinomas (5,6). This antibody has been referred to as "RD20" by Peng et al.(1).

References

  1. Peng et al. (2013) Cell Rep. 3(6):1885.
  2. de Lau et al. (2014) Genes Dev. 28(4):305.
  3. Barker et al. (2013) Development 140(12):2484.
  4. Clevers (2013) Nature 495(7439):53.
  5. Wu et al. (2014) Nat Commun 5:3149.
  6. Jang et al. (2013) PLoS One 8(12):e82390495.

Long Name

Leucine-rich Repeat Containing G Protein-coupled Receptor 5

Alternate Names

FEX, GPR49, GPR67, HG38

Entrez Gene IDs

8549 (Human); 14160 (Mouse); 299802 (Rat); 102120216 (Cynomolgus Monkey)

Gene Symbol

LGR5

UniProt

Q9Z1P4

Additional Lgr5/GPR49 Products

Product Documents for Mouse Lgr5/GPR49 Antibody (803420)

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Product Specific Notices for Mouse Lgr5/GPR49 Antibody (803420)

For research use only

Citations for Mouse Lgr5/GPR49 Antibody (803420)

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