Human Lgr4/GPR48 Antibody

Catalog #: MAB7750 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB7750
MAB7750-SP

Detection of Lgr4/GPR48 in HEK293 Human Cell Line Transfected with Human Lgr4/GPR48 and eGFP by Flow Cytometry.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Lgr4/GPR48 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Lgr4/GPR48 in Lgr4-transfected cell-based ELISA. Stains human Lgr4/GPR48 transfected but not irrelevant transfected cells in flow cytometry.

Source

Monoclonal Mouse IgG_2B Clone # 852229

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

NS0 mouse myeloma cell line transfected with human Lgr4/GPR48
Met1-Asp951
Accession # Q9BXB1

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

Blockade of Receptor-ligand Interaction

In a functional flow cytometry test, 2.5 μg/mL of Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody completely blocks binding of biotinylated Recombinant Human R-Spondin 4 (100 ng/mL) to HEK293 human embryonic kidney cell line transfected with human Lgr4/GPR48.

CyTOF-ready

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

Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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Detection of Lgr4/GPR48 in HEK293 Human Cell Line Transfected with Human Lgr4/GPR48 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with human Lgr4/GPR48 and eGFP was stained with and either (A) Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody (Catalog # MAB7750) or (B) Mouse IgG_2BFlow Cytometry Isotype Control (Catalog # MAB0041) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry

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Lgr4/GPR48 in HT‑29 Human Cell Line. Lgr4/GPR48 was detected in immersion fixed HT-29 human colon adenocarcinoma cell line using Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody (Catalog # MAB7750) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Blockade of Receptor-ligand Interaction

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R-Spondin 4 Binding to Lgr4/GPR48-transfected HEK293 Human Cell Line is Blocked by Human Lgr4/GPR48 Antibody. In a functional flow cytometry test, biotinylated Recombinant Human R-Spondin 4 (100 ng/mL) binds to HEK293 human embryonic kidney cell line transfected with human Lgr4/GPR48 (dark orange histogram). Binding is completely blocked (light orange histogram) by 2.5 µg/mL of Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody (Catalog # MAB7750). Mouse IgG2B Isotype Control (Catalog # MAB004) at 2.5 µg/mL was used as a control (blue line). Cells were stained with Streptavidin-APC (Catalog # F0050).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

Background: Lgr4/GPR48

Lgr4 (Leucine-rich repeat GPR 4), also known as GPR48 (G-Protein-coupled Receptor 48), is a seven-transmembrane glycoprotein receptor in the Lgr family of cell surface receptors (1, 2). While this family includes receptors for hormones such as LH, FSH, TSH, and HCG, the subfamily comprising Lgr4, Lgr5, and Lgr6 are G-protein-independent mediators of the potentiating effect of R-Spondins on Wnt signaling (1-6). Lgr4 binds and forms complexes with R-Spondins, Frizzled Wnt receptors and LRP Wnt co-receptors (5). It acts at least in part by enhancing Wnt-dependent LRP phosphorylation, internalization of LRPs, and accumulation of beta -catenin (3, 4). Human Lgr4 cDNA encodes 951 amino acids (aa), including a long N-terminal Extracellular Domain (ECD, aa 25-544) with 16-17 LRR domains that mediate ligand interaction (1). The LRR-containing ECD of human Lgr4 shares 93% aa sequence identity with mouse, rat and bovine Lgr4, and 50-60% aa identity with human Lgr5 and Lgr6. Lgr4 is widely expressed in both embryo and adult. Expression of Lgr4 mRNA in adult humans is highest in pancreas, followed by liver, heart, muscle, brain, and placenta (1). In rodents, embryonic and adult expression includes liver, kidney, adrenals, bone/cartilage, and heart (2, 7-9). Lgr4 deletion in the mouse affects development in areas of expression, for example, inhibiting fetal liver definitive erythropoiesis (9). Deletion of Lgr4 specifically from stem and progenitor cells in intestinal crypts induces loss of crypts due to insufficient Wnt signaling (5, 6). Lgr4 may be over-expressed in carcinomas and may promote invasiveness and metastasis by down-regulating p27^Kip1 expression (10).

References