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Safety Data Sheet (SDS) Purified Anti-mouse CD100 SEMA4D Antibody     Product Data Sheet (PDF)    
Purified Anti-mouse CD100 (SEMA4D) Antibody
1338005 25 µg $80.00       
1338010 100 µg $260.00       
Clone: BMA-12
Isotype: Rat IgG2a, κ
Reactivity: Mouse
Immunogen: Mouse CD100-Fc fusion protein
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application:

FC - Quality tested
IHC-F - Validated
IP, ELISA - Reported in the literature

Application Notes:

Additional reported applications for the relevant formats include: immunoprecipitation from cell lysates2, capture antibody function in a sandwich ELISA assay2, and immunohistochemical staining of frozen brain sections4.

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. For immunohistochemical staining on frozen tissue sections, the suggested use of this reagent is 5.0 - 10 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application References:

1. Kumanogoh A, et al. 2000. Immunity. 13:621. (FC)
2. Wang X, et al. 2001. Blood. 97:3498. (FC, IP, ELISA)
3. Kumanogoh A, et al. 2002. J. Immunol. 169:1175. (FC)
4. Moreau-Fauvarque C, et al. 2003 J. Neurosci. 23:9229. (IHC)

C57BL/6 mouse splenocytes were stained

C57BL/6 mouse splenocytes were stained with purified CD100/SEMA4D (clone BMA-12, filled histogram) or purified rat IgG2a, κ isotype control (open histogram), followed by anti-rat IgG PE.

C57BL/6 mouse frozen kidney section

C57BL/6 mouse frozen kidney section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then the section was stained with 10 µg/mL of purified anti-mouse CD100 (clone BMA-12) and anti-mouse Podoplanin (clone 8.1.1) Alexa Fluor® 594 (green) overnight at 4°C, followed by 2.5 µg/mL of goat anti-mouse IgG (clone Poly4053) Alexa Fluor® 647 (red) for two hours at room temperature. The image was captured with a 10X objective.



Description:

CD100, also known as Semaphorin-4D (SEMA4D), is a 150 kD, type I transmembrane protein, with one sema domain and one C2-type Ig domain. It is found as a homodimer on the cell surface. CD100 is expressed in the brain, kidney, heart, the majority of hematopoietic cells, and some malignant cells. CD100 is a potent inducer of angiogenesis and inhibitor of bone formation, and plays a role in lymphocyte activation, antibody production, dendritic cell maturation, and axon guidance. Its ligands are CD72 and Plexin-B1. CD100 is cleaved from the membrane of activated T and B cells, producing a soluble form.

Other Names: Neuroimmune semaphorin 4D, Sema4D, M-Sema G, Semaphorin-C-like 2, Semaphorin-J, Semaj, Semcl2, Coll-4, Semacl2
Structure: Type I transmembrane protein, one sema domain, one C2-type immunoglobulin domain, homodimer, 150 kD.
Distribution: Highly expressed on most hematopoietic cells, is also found in the brain, kidney, and heart. Overexpressed by some malignant cell types.
Function: Potent inducer of angiogenesis, inhibitor of bone formation, plays a role in lymphocyte activation, antibody production and dendritic cell maturation. In neurons, regulates dendrite branching and axon guidance.
Ligand Receptor: CD72, Plexin-B1.
Antigen References:

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell. 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.