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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 (MA3218) at 2µg/ml 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 (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 (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 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.
|Tested species reactivity||Human, Pig|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified extracellular domain of recombinant human thyrotropin receptor.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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.
The thyroid stimulating hormone (thyrotropin) receptor (TSHR) is expressed on the surface of thyrocyte cell membranes and is crucial for thyroid gland growth and function. TSHR"e;s are targets for the pathogenesis of many autoimmune diseases including Graves"e; disease and primary myxedema. This 7-transmembrane-domain protein contains 5 potential extracellular N-linked glycosylation sites. Antibodies to TSHR can affect receptor function in a number of different ways. TSHR antibodies which can inhibit TSH binding to the receptor are called TSH binding inhibitor immunoglobulins or TBIIs.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
seven transmembrane helix receptor; thyroid stimulating hormone receptor, isoform 2; thyroid-stimulating hormone receptor; thyrotropin receptor; thyrotropin receptor-I, hTSHR-I; TSH-R; TSHR
CHNG1; hTSHR-I; LGR3; TSHR