Mouse IgG1 kappa Isotype Control [Brilliant Violet 785] (DAGIC614)

Mouse IgG1 kappa Isotype Control for FC, ICFC

Additional Formats Available

Specifications


Host Species
Mouse
Antibody Isotype
IgG1, κ
Clone
NPQD-32
Species Reactivity
N/A
Conjugate
Brilliant Violet 785

Applications


Application Notes
FC, ICFC
*Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.

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References


A human monoclonal autoantibody to the thyrotropin receptor with thyroid-stimulating blocking activity

THYROID

Authors: Sanders, Jane; Evans, Michele; Betterle, Corrado; Sanders, Paul; Bhardwaja, Anshu; Young, Stuart; Roberts, Emma; Wilmot, Jane; Richards, Tonya; Kiddie, Angela; Small, Kasemsri; Platt, Hayley; Summerhayes, Sara; Harris, Rebecca; Reeve, Magnus; Coco, Graziella; Zanchetta, Renato; Chen, Shu; Furmaniak, Jadwiga; Smith, Bernard Rees

Background: Human monoclonal autoantibodies (MAbs) are valuable tools to study autoimmune responses. To date only one human MAb to the thyrotropin (TSH) receptor (TSHR) with stimulating activity has been available. We now describe the detailed characterization of a blocking type human MAb to the TSHR. Methods: A single heterohybridoma cell line was isolated from the peripheral blood lymphocytes of a patient with severe hypothyroidism (TSH 278 mU/L) using standard techniques. The line stably expresses a TSHR autoantibody (5C9; IgG1/kappa). Ability of 5C9 to bind and compete with I-125-TSH or TSHR antibodies binding to the TSHR was tested using tubes coated with solubilized TSHR. Furthermore, the blocking effects of 5C9 on stimulation of cyclic AMP production was assessed using Chinese hamster ovary (CHO) cells expressing the wild-type human TSHR or TSHRs with amino acid mutations. Main outcome: 5C9 IgG bound to the TSHR with high affinity (4 x 10(10) L/mol) and inhibited binding of TSH and a thyroid-stimulating human monoclonal autoantibody (M22) to the receptor. 5C9 IgG preparations inhibited the cyclic AMP-stimulating activities of TSH, M22, serum TSHR autoantibodies and thyroid-stimulating mouse monoclonal antibodies. Furthermore 5C9 reduced the constitutive activity of wild-type TSHR and TSHR with some activating mutations. The effect of different amino acid mutations in the TSHR on 5C9 biological activity was studied and TSHR Lys129Ala or Asp203Ala completely abolished the ability of 5C9 to block TSH-mediated stimulation of cyclic AMP production. Conclusions: The availability of 5C9 provides new opportunities to investigate the binding and biological activity of TSHR blocking type autoantibodies including studies at the molecular level. Furthermore, monoclonal antibodies such as 5C9 may well provide the basis of new drugs to control TSHR activity including applications in thyroid cancer and Graves' ophthalmopathy.

A Novel Immunoglobulin-Immunoglobulin Interaction in Autoimmunity

PLOS ONE

Authors: Kawa, Shigeyuki; Kitahara, Kei; Hamano, Hideaki; Ozaki, Yayoi; Arakura, Norikazu; Yoshizawa, Kaname; Umemura, Takeji; Ota, Masao; Mizoguchi, Sadaaki; Shimozuru, Yasunori; Bahram, Seiamak

Well over six decades since its first description, the Rheumatoid Factor (RF)-autoantibodies recognizing Fc (constant) portion of IgG through their own Fab (antigen binding variable segments)-is believed to have come of age. Autoimmune pancreatitis is a unique form of pancreatitis, biologically characterized by an elevated serum IgG4 concentration. Given the fact that IgG4 myeloma proteins can act as RF, we initially hypothesized that IgG4 in autoimmune pancreatitis might do likewise, hence potentially contributing to disease pathogenesis. Indeed Western blotting clearly showed that IgG4 binds to IgG1 kappa, IgG2 kappa, IgG3 kappa myeloma proteins, as well as to IgG Fc, in line with a typical RF activity. Further experiments however unraveled the unexpected fact that unlike hitherto known RF, IgG4 does not engage IgG Fc through its Fab, but its very own Fc. These data therefore collectively describe a Novel RF (NRF) in autoimmune pancreatitis. In the future, the relevance of NRF, beyond autoimmune pancreatitis, in both diagnosis/prognosis as well as pathophysiology of autoimmune and other systemic diseases where IgG4's role seems paramount, needs to be systematically assessed.

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