Sunyer Laboratory

CLONE NAME: 41.8                    
HOST/ISOTYPE: MOUSE/IgG2b

BACKGROUND: Three immunoglobulin classes (IgM, IgD and IgT) are present in teleost fish.  IgT, also called IgZ in some fish species, was identified in 2005. In rainbow trout, there are two major B cell subsets expressing either IgT or IgM/IgD, but no B cells expressing both IgM and IgT.  In contrast to IgM+ B cells, IgT+  B cells do no co-express IgD. Significant titers of rainbow trout IgT and accumulation ofs IgT+ B cells are detected in mucus and mucosal tissues (skin and gut) respectively from fish infected with mucosal pathogens. Moreover, bacterial microbiota in mucus from gut and skin are prevalently coated by IgT.  Therefore, IgT and IgT+ B cells play specialized roles in rainbow trout mucosal immune responses and homeostasis. The above information regarding the production of IgT monoclonal antibodies and the detection of IgT and IgT+  B cells in rainbow trout gut and skin have been reported in the publications shown in the “references” section below.

SPECIFICITY:  The clone  41.8 produces mouse IgG2b that recognizes refolded recombinant protein of the constant domains 2-4 of rainbow trout IgT heavy chain which was recombinantly produced in E. coli. This monoclonal antibody recognizes soluble  IgT from rainbow trout fluids (plasma, serum, skin mucus, gut mucus) as well as membrane IgT from  IgT+ B cells. The antibody specificity was demonstrated in publication #1 of the ”references” section. 

IMMUNOGEN: Recombinant protein of heavy chain constant domain (CH2-4 domains) of rainbow trout produced in E. coli.

APPLICATIONS: 

• Western blot: 2 μg/ml. Recognizes IgT from serum, plasma, whole cell lysates.

• Perform SDS-PAGE under non-reducing conditions prior to blotting proteins.
• FACS: 2 μg/ml for 100 μL cell (1×107 cells/mL)

 PRESENTATION: Protein G purified immunoglobulin in PBS.

STORAGE/HANDLING: Store antibody in aliquot, tightly sealed at +2-4 oC, although sodium azide or Bronidox (both at 0.05%) is recommended as preservative if stored at that temperature for more than 1 day. It is important to avoid multiple thaw and freezing cycles as well as warm and chill cycles. If product requires storage for longer than 1-2 weeks, it is recommended that it is stored at -20-80 oC.

REFERENCES:

1. Zhang, Y. A., I. Salinas, J. Li, D. Parra, S. Bjork, Z. Xu, S. E. LaPatra, J. Bartholomew, and J. O. Sunyer. 2010. IgT, a primitive immunoglobulin class specialized in mucosal immunity. Nat. Immunol. 11:827-835.
2. Xu, Z., Parra, D., Gomez, D., Salinas, I., Zhang, Y.A., Jørgensen G.L., Skjødt, K., Rasmussen, K.J.,Buchmann K., LaPatra, S.L., J. O. Sunyer. 2013.  Teleost skin, an ancient mucosal surface that elicits gut-like immune-responses. PNAS.110:13097-13102.

Important notification: It is possible that during shipment, small volumes of product may become entrapped in the seal of the enclosed vial. Therefore, for products with 250 μl or less, we suggest centrifuging the vial in a tabletop centrifuge in order to dislodge any product in the container’s cap.

HOST/ISOTYPE: Rabbit/IgG

BACKGROUND: Three immunoglobulin classes (IgM, IgD and IgT) are present in teleost fish.  IgT, also called IgZ in some fish species, was identified in 2005. In rainbow trout, there are two major B cell subsets expressing either IgT or IgM/IgD, but no B cells expressing both IgM and IgT.  In contrast to IgM+ B cells, IgT+  B cells do no co-express IgD. Significant titers of rainbow trout IgT and accumulation ofs IgT+ B cells are detected in mucus and mucosal tissues (skin and gut) respectively from fish infected with mucosal pathogens. Moreover, bacterial microbiota in mucus from gut and skin are prevalently coated by IgT.  Therefore, IgT and IgT+ B cells play specialized roles in rainbow trout mucosal immune responses and homeostasis. The above information regarding the production of IgT polyclonal antibodies and the detection of IgT and IgT+  B cells in rainbow trout gut and skin have been reported in the publications shown in the “references” section below.

SPECIFICITY:   This polyclonal antibody recognizes the refolded recombinant protein of the constant domains 2-4 of rainbow trout IgT heavy chain which was recombinantly produced in E. coli. This polyclonal antibody recognizes soluble  IgT from rainbow trout fluids (plasma, serum, skin mucus, gut mucus) as well as membrane IgT from  IgT+ B cells. The antibody specificity was demonstrated in publication #1 of the ”references” section.

IMMUNOGEN: Recombinant protein of heavy chain constant domain (CH2-4 domains) of rainbow trout produced in E. coli.

APPLICATIONS: 

• Western blot: 1 μg/ml. Recognizes IgT from serum, plasma, whole cell lysates.
• Perform SDS-PAGE under reducing or non-reducing conditions prior to blotting proteins.
• FACS: 2 μg/ml for 100 μL cell (1×107 cells/mL)

PRESENTATION: Rabbit IgG was first purified by protein G chromatography. Thereafter, specific IgG was affinity purified against a column containing the recombinant immunogen. The antibody is in PBS.

STORAGE/HANDLING: Store antibody in aliquot, tightly sealed at +2-4 oC, although sodium azide or Bronidox (both at 0.05%) is recommended as preservative if stored at that temperature for more than 1 day. It is important to avoid multiple thaw and freezing cycles as well as warm and chill cycles. If product requires storage for longer than 1-2 weeks, it is recommended that it is stored at -20-80 oC.

REFERENCES:

1. Zhang, Y. A., I. Salinas, J. Li, D. Parra, S. Bjork, Z. Xu, S. E. LaPatra, J. Bartholomew, and J. O. Sunyer. 2010. IgT, a primitive immunoglobulin class specialized in mucosal immunity. Nat. Immunol. 11:827-835.
2. Xu, Z., Parra, D., Gomez, D., Salinas, I., Zhang, Y.A., Jørgensen G.L., Skjødt, K., Rasmussen, K.J.,Buchmann K., LaPatra, S.L., J. O. Sunyer. 2013.  Teleost skin, an ancient mucosal surface that elicits gut-like immune-responses. PNAS.110:13097-13102.

Important notification: It is possible that during shipment, small volumes of product may become entrapped in the seal of the enclosed vial. Therefore, for products with 250 μl or less, we suggest centrifuging the vial in a tabletop centrifuge in order to dislodge any product in the container’s cap.