Invitrogen

TSH Receptor Monoclonal Antibody (49)

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TSH Receptor Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Thyrotropin Receptor was performed using 70% confluent log phase A431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with TSH Receptor (49) Mouse Monoclonal Antibody (Product # MA3-218) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

TSH Receptor Antibody in Western Blot (WB) — Western blot analysis was performed on membrane-enriched extracts (30 µg lysate) of A-431 (Lane 1), IMR-32 (Lane 2) and Jurkat (Lane 3). The blots were probed with Anti-TSH Receptor Mouse Monoclonal Antibody (Product # MA3-218, 1:250 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 63 kDa band corresponding to TSH Receptor was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Product Details

MA3-218

Applications	Tested Dilution	Publications
Western Blot (WB)	1:250	-
Immunocytochemistry (ICC/IF)	1:250	-
ELISA (ELISA)	Assay-dependent	-
Miscellaneous PubMed (Misc)	-	View 3 publications 3 publications

Product Specifications
Species Reactivity	Human, Pig
Published species	Not Applicable
Host/Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	49
Immunogen	Purified extracellular domain of recombinant human thyrotropin receptor.
Conjugate	Unconjugated
Form	Liquid
Concentration	Conc. Not Determined
Storage buffer	ascites
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_325488

Product Specific Information

MA3-218 detects both glycosylated and nonglycosylated thyroid stimulating hormone receptor (TSHR) from porcine tissues as well as recombinant human TSHR.

MA3-218 has been successfully used in Western blot, immunofluorescence, and ELISA procedures. By Western blot, this antibody detects a 50 kDa and a 63 kDa protein representing unglycosylated and glycosylated TSHR respectively, in transfected Sf9 cell extract. By ELISA MA3-218 has been used in TSH competitive inhibition studies.

The MA3-218 antigen is purified extracellular domain of recombinant human thyrotropin receptor. This antibody recognizes an epitope between amino acid residues 277-296 of human TSHR.

Target Information

TSH Receptor gene is a membrane protein and a major controller of thyroid cell metabolism. The encoded protein is a receptor for thyrothropin and thyrostimulin, and its activity is mediated by adenylate cyclase. Defects in this gene are a cause of several types of hyperthyroidism. Three transcript variants encoding different isoforms have been found for this gene.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

The thyrotropin (thyroid-stimulating hormone) receptor is expressed on murine dendritic cells and on a subset of CD45RBhigh lymph node T cells: functional role for thyroid-stimulating hormone during immune activation.

Journal of immunology (Baltimore, Md. : 1950)

Bağriaçik EU,Klein JR

MA3-218 was used in western blot and immunoprecipitation to study the expression and function of thyrotropin receptor in murine dendritic cells and in lymph node T cells .

Thu Jun 15 00:00:00 EDT 2000

Differential reactivities of recombinant glycosylated ectodomains of mouse and human thyrotropin receptors with patient autoantibodies.

Endocrinology

Patibandla SA,Seetharamaiah GS,Dallas JS,Thotakura NR,Peake RL,Prabhakar BS

MA3-218 was used in ELISA to investigate the expression of mET-gp and characterize the difference of epitopes between mET-gp and ETSHr-gp

Tue Apr 01 00:00:00 EST 1997

Generation and characterization of monoclonal antibodies to the human thyrotropin (TSH) receptor: antibodies can bind to discrete conformational or linear epitopes and block TSH binding.

Endocrinology

Seetharamaiah GS,Wagle NM,Morris JC,Prabhakar BS

MA3-218 was used in blocking/activating experiment and ELISA to characterize human TSH receptor antibodies in vitro and in vivo

Sat Jul 01 00:00:00 EDT 1995

Bioinformatics

Protein Aliases: MGC141920; seven transmembrane helix receptor; thyroid stimulating hormone receptor, isoform 2; Thyroid-stimulating hormone receptor; Thyrotropin receptor; thyrotropin receptor-I, hTSHR-I; TSH-R; TSHR; seven transmembrane domain G protein-coupled receptor

Gene Aliases: CHNG1; hTSHR-I; LGR3; TSHR

UniProt ID: (Pig) Q8SPP9, (Human) P16473

Entrez Gene ID: (Pig) 397560, (Human) 7253

Molecular Function: G-protein coupled receptor