Immunochemistry

Research for Immune System

Anti Mouse Sema4A, MAb (1H9)
Anti Mouse Sema4D/CD100, MAb (5H7)

Anti Sema4A (Wako Catalog No. 019-22811 (100 μL); Anti Sema4D (Wako Catalog No. 016-22821 (100 μL)

The expression of semaphorins are seen most fully in the nervous system, and also present in most other tissues. Functionally, semaphorins were initially characterized for their importance in the development of the nervous system and in axonal guidance.  Lately, they have been found to be important for the formation and functioning of neural connectivity, angiogenesis, immunoregulation, and cancer.

Outlines

Sema4A is expressed at high levels in dentritic cells in immune system.  Although no expression is in dormant T cells, the expression of sema4A is induced when T cells is differentiated into Th1 type cells.
Furthermore, an immunological analysis has found that Sema4A relates to Th1/Th2 control.

 Sema4D/CD100, which belongs to the semaphorin family, is expressed at high levels in T cellls.  Although the expression is weak in dormant B cells and dendritic cells, the increased expression are in those activated cells.  It also plays an important roles in immune response and in maintaining immune homeostasis.

Product Profile

Anti Mouse Sema4A, MAb (1H9)

Appearance: the PBS solution
Concentration: 0.7 mg/mL
    (an actual measured value of the first lot)
Specificity: mouse and human Sema4A
Recommended Working Dilution:
    Immunocytochemistry  1 : 20 ~ 1 : 50
    Immunohistochemistry  1 : 10 ~ 1 : 20
    Immunoprecipitation    1 : 500 ~ 1 : 1,000

Anti Mouse Sema4D/CD100, MAb (5H7)

Appearance: the PBS solution
Concentration: 0.9 mg/mL
    (an actual measured value of the first lot)
Specificity: mouse and human Sema4D/CD100
Recommended Working Dilution:
  Immunocytochemistry  1 : 20 ~ 1 : 50
  Immunoprecipitation    1 : 100 ~ 1 : 500

Applications

Immunoprecipitation (IP)
Sema4A-FLAG (DYKDDDDK) (abt. 110 k) and Sema4D/CD100-FLAG (abt. 150 k) were forcedly expressed in COS7 cells and the cells were lysed by NP-40. After immunoprecipitation with Anti Mouse Sema4A and Anti Mouse Sema4D (5 μg/mL lysate), respectively, SDS-PAGE was performed and Western blot was carried out using Anti DYKDDDDK Antibody. Anti Mouse IgG-POD was used as a second antibody and ImmunoStarR LD was used for detection.

PHOTO

FACS
Sema4A and Sema4D were expressed in COS7 cells. The cells were stained with Anti Mouse Sema4A and Anti Mouse Sema4D (1:30), with Anti Mouse IgG (1:50) used as a second antibody, and detected by Allophycocyanin.

PHOTO
Green line: control; Red line: COS7 cells, in which Sema4A or Sema4D/CD100 was expressed
Data was provided by Kumanogoh, A., Dept. of Molecular Immunology, Research Institute for Microbial Diseases, Osaka Univ.

References:
1.  "Nonredundant roles of Sema4A in the immune system: defective T cell priming and Th1/Th2 regulation in Sema4A-deficient mice", Kumanogoh, A. et al.: Immunity, 22, 305-16 (2005)
2.  "Roles of Sema4D-Plexin-B1 Interactions in the Central Nervous System for Pathogenesis of Experimental Autoimmune Encephalomyelitis", Okuno, T. et al.: J. Immunol. 184, 1499-1506 (2010)