The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 4 - 6 µg/ml.
Members of the semaphorin protein family are involved in axonal guidance during neural development. Semaphorin 5a is thought to have a bifunctional role in axon guidance, exerting both attractive and inhibitory effects on developing axons associated with limbic function. The thrombospondin repeats of Semaphorin 5a physically interact with both heparan sulfate proteoglycans and chondroitin sulfate proteoglycans, inducing attractive and inhibitory guidance cues, respectively. The biological responses to Semaphorin 5a are induced through binding to its high-affinity receptor, plexin B3, triggering intracellular signaling through Met.
Immunohistochemistry (Frozen sections) - Anti-Semaphorin 5A antibody (ab51957)This image is courtesy of an Abreview submitted by Mrs Elena Astanina
ab51957 (1/200) detecting Semaphorin 5A in Chicken E11 embryo. Tissue was fixed in paraformaldehyde, permeabilised with 0.2% Triton X-100/PBS and blocked in 5% serum for 1 hour at 37oC. Incubation with primary antibody was performed for 16 hours in 0,5% BSA/ 2,5% serum/ PBS at 4oC. Please refer to abreview for further experimental details.
ab51957 at 4µg/ml staining Semaphorin 5A in Human kidney tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Steamed antigen retrieval with citrate buffer pH 6 was performed. Antibody was detected by HRP-staining.
Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 5A antibody (ab51957)Image from Monticone M et al., BMC Cancer. 2012 Aug 17;12:358. doi: 10.1186/1471-2407-12-358. Fig 6.; 17 August 2012, BMC Cancer 2012, 12:358
Immunofluorescence analysis of Human Glioblastoma multiforme cells, staining Semaphorin 5A with ab51957.
Cells were fixed in 3% paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with pure goat serum for 30?min on ice. Samples were incubated with primary antibody for 2 hours on ice. An AlexaFluor®488-conjugated anti-goat IgG was used as the secondary antibody.