Anti-Semaphorin 4A Antibody

• Catalog Number: orb215259
• Category: Antibodies
• Description: Rabbit polyclonal antibody to SEMA4A
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IF, IH, IP, WB
• Reactivity: Human, Primate
• Immunogen: KLH-conjugated synthetic peptide encompassing a sequence within the center region of human Semaphorin 4A. The exact sequence is proprietary.
• Antibody Type: Primary Antibody
• Dilution range: WB: 1-500-1-1000, IF/ICC: 1-100-1-500, IP: 1-10-1-100
• Form/Appearance: Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
• Conjugation: Unconjugated
• Target: SEMA4A
• Entrez: 64218
• UniProt ID: Q9H3S1
• Source: Rabbit
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
• Alternative names: anti SEMAB antibody, anti SEMB antibody, anti Sema
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Semaphorin 4A expression in HEK293T (A), A549 (B), SGC7901 (C) whole cell lysates. (Predicted band size: 83 kD; Observed band size: 95 kD)

Immunohistochemical analysis of Semaphorin 4A staining in human brain formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of Semaphorin 4A staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunoprecipitation of Semaphorin 4A from 0.5 mg Jurkat whole cell extract lysate, using 5 ug of Anti-Semaphorin 4A Antibody and 50 ul of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10 min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 min under agitation. Proteins were eluted by addition of 40 ul SDS loading buffer and incubated for 10 min at 70°C; 10 ul of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with Anti-Semaphorin 4A Antibody.