Mouse Anti-Semaphorin 4B (SEMA4B, Semaphorin-4B, KIAA1745, SEMAC, UNQ749/PRO1480)
Semaphorin 4B (Sema4B; also SemaC) is a 95-100kD, class IV member of the semaphorin family of proteins. It is expressed in neurons, and following PSD-95 induced clustering, participates in the formation or functioning of glutamatergic synapses. Mature human Semaphorin 4B is a type I transmembrane glycoprotein that contains one Sema domain (aa65-502), a PSI region (aa520-574), and an Ig-like C2-type domain (aa599-658) in its extracellular domain. There is one potential soluble splice variant that shows a 29aa substitution for aa707-832. Over aa39-712, human Semaphorin 4B shares 86% aa identity with mouse Semaphorin 4B.
Applications
Suitable for use in Flow Cytometry and Direct ELISA. Other applications not tested.
Recommended Dilutions
Flow Cytometry: 0.25ug/10e6 cells using THP-1 human acute monocytic leukemia cell line Optimal dilutions to be determined by the researcher.
Hybridoma
Fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, NS0-derived, recombinant human Semaphorin 4B.
Flow Cytometry Protocol
1. This antibody was tested for flow cytometry using THP-1 cells. 2. Dilute S0668-55 to 25ug/ml and add 10ul of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200ul. 3. Wash cells with F4568-10 Flow Cytometry Staining Buffer (1X). 3. The binding of unlabeled antibodies may be visualized by adding a secondary developing reagent such as anti-mouse IgG conjugated to a fluorochrome.
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
Recombinant protein corresponding to Leu39-Glu712 of human Semaphorin 4B, expressed in NS0 cells.
Form
Supplied as a lyophilized powder from PBS, pH 7.4, 5% trehalose. Reconstitute with 200ul sterile PBS.
Purity
Purified by Protein G affinity chromatography from hybridoma culture supernatant.
Specificity
Recognizes human Semaphorin 4B. Does not react with mouse Semaphorin 4B.