Overview

  • Product name
    Anti-TSH Receptor/TSH-R antibody [A7]
    See all TSH Receptor/TSH-R primary antibodies
  • Description
    Mouse monoclonal [A7] to TSH Receptor/TSH-R
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Fusion protein corresponding to Human TSH Receptor/TSH-R aa 402-415 (C terminal).

  • Epitope
    The murine monoclonal antibody A7 is specific for residues 402-415 of the human TSH receptor. This epitope is localized at the extreme carboxyl terminal of the extracellular domain of the TSH receptor, a region that may be masked from the surface of native TSH receptor.
  • Positive control
    • ICC/IF: HEK-293 cells. Flow Cyt: HeLa cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.4
    Constituents: PBS, 0.81% Sodium chloride, 0.16% Sodium phosphate, 0.02% Potassium chloride, 0.04% Potassium phosphate
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    A7
  • Isotype
    IgG2b
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6044 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Receptor for thyrothropin. Plays a central role in controlling thyroid cell metabolism. The activity of this receptor is mediated by G proteins which activate adenylate cyclase. Also acts as a receptor for thyrostimulin (GPA2+GPB5).
  • Tissue specificity
    Expressed in the thyroid.
  • Involvement in disease
    Defects in TSHR are found in patients affected by hyperthyroidism with different etiologies. Somatic, constitutively activating TSHR mutations and/or constitutively activating G(s)alpha mutations have been identified in toxic thyroid nodules (TTNs) that are the predominant cause of hyperthyroidism in iodine deficient areas. These mutations lead to TSH independent activation of the cAMP cascade resulting in thyroid growth and hormone production. TSHR mutations are found in autonomously functioning thyroid nodules (AFTN), toxic multinodular goiter (TMNG) and hyperfunctioning thyroid adenomas (HTA). TMNG encompasses a spectrum of different clinical entities, ranging from a single hyperfunctioning nodule within an enlarged thyroid, to multiple hyperfunctioning areas scattered throughout the gland. HTA are discrete encapsulated neoplasms characterized by TSH-independent autonomous growth, hypersecretion of thyroid hormones, and TSH suppression. Defects in TSHR are also a cause of thyroid neoplasms (papillary and follicular cancers).
    Autoantibodies against TSHR are directly responsible for the pathogenesis and hyperthyroidism of Graves disease. Antibody interaction with TSHR results in an uncontrolled receptor stimulation.
    Hypothyroidism, congenital, non-goitrous, 1
    Familial gestational hyperthyroidism
    Hyperthyroidism, non-autoimmune
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family. FSH/LSH/TSH subfamily.
    Contains 7 LRR (leucine-rich) repeats.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CHNG1 antibody
    • hTSHR I antibody
    • hTSHRI antibody
    • LGR 3 antibody
    • LGR3 antibody
    • MGC75129 antibody
    • Seven transmembrane helix receptor antibody
    • Thyroid adenoma hyperfunctioning antibody
    • Thyroid carcinoma with thyrotoxicosis antibody
    • Thyroid stimulating hormone receptor antibody
    • Thyroid stimulating hormone receptor, isoform 2 antibody
    • Thyroid-stimulating hormone receptor antibody
    • Thyrotropin receptor antibody
    • Thyrotropin receptor I antibody
    • Thyrotropin receptor I, hTSHR I antibody
    • TSH R antibody
    • TSH Receptor antibody
    • TSH-R antibody
    • TSHR antibody
    • TSHR_HUMAN antibody
    see all

Images

  • ICC/IF image of ab6044 stained Hek293 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6044, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • Overlay histogram showing HeLa cells stained with ab6044 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6044, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91633, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Arauchi A  et al. Functional Thyroid Follicular Cells Differentiation from Human-Induced Pluripotent Stem Cells in Suspension Culture. Front Endocrinol (Lausanne) 8:103 (2017). Read more (PubMed: 28588551) »
  • Dorris ER  et al. Pluripotency markers are differentially induced by MEK inhibition in thyroid and melanoma BRAFV600E cell lines. Cancer Biol Ther 17:526-42 (2016). WB . Read more (PubMed: 26828826) »
See all 4 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question

Please read below customer's answers:

1. To explain both of the western blot images? Conditions used in each blot? Conditions for each lane?

24.9.12 – The big membrane was divided into four membranes according to size of the desirable proteins. On both sides the same amount of protein was loaded (and it is the same origin – human thyroid cells). The two lower membranes are control for loading. The two sides are duplicates. The difference is the antibody used: left: ab-TSHR right: ab-MMP9. The conditions used for each lane were identical.

The following is related to all four membranes in the image 24.9.12 and for the two membranes in the image 11.10.12:

Lane 1: marker

Lane 2: normal thyroid 15µg

Lane 3: normal thyroid 40µg

lane 4: thyroid tumor 15µg

Lane 5: thyroid tumor 40µg

Lane 6: thyroid metastasis 15µg

Lane 7: thyroid metastasis 40µg

11.10.12 The two membranes were stripped and I switched the membranes and reacted the left one with ab-MMP9 and the right with ab-TSHR in order to check if the problem was because of the specific membrane but again I got high background with no clue for the right bands. In this time I also diluted the ab two times more than recommended and diluted the ECL two times more as detailed in the complaint form.

Each membrane was reacted with ECL according to the manufacturer's instructions.


2. For image labeled 24/09/12, why is the top half of the blot overexposed when the bottom half isn't? Was this blot cut in half and treated differently? Exposure time was the same for all four membranes – about 1 second. The longer exposure times gave even darker images. The four membranes were treated in the same way and were exposed for the same time. The differences between the four membranes are only the antibodies.

3. How long was this membrane exposed for? For 1-2 seconds.

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.
I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.
Unfortunately, if you would like a free of charge replacement, we don't have any other lot in stock and I am reluctant to send you the same lot again as I fear that you may get the same results. I can recommend ab27974 (www.abcam.com/ab27974) as an alternative.
Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.
Could you ask your customer to clarify the following:
1. To explain both of the western blot images? Conditions used in each blot? Conditions for each lane?
2. For image labelled 24/09/12, why is the top half of the blot overexposed when the bottom half isn't? Was this blot cut in half and treated differently?
3. How long was this membrane exposed for?
In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.
I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you for your inquiry. I can confirm that this antibody is sold in the following buffer: PBS (0.14 M Sodium Chloride; 0.003 M Potassium Chloride; 0.002 M Potassium Phosphate; 0.01 M Sodium Phosphate; pH 7.4) with  no preservative or other additives. I hope this information was helpful. Please do not hesitate to contact me again with any further questions.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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