CD3 antibody

• Catalog Number: orb44679
• Category: Antibodies
• Description: Mouse monoclonal antibody to CD3
• Clonality: Monoclonal
• Clone Number: APA1/1
• Tested applications: FC, ICC, IHC-Fr, IP, WB
• Reactivity: Human, Mouse
• Isotype: Mouse IgG1
• Immunogen: Purified human CD3 proteins isolated from thymus
• Concentration: 1 mg/ml
• Dilution range: Flow cytometry: Intracellular staining; recommended dilution: 1-4 µg/ml; positive control: human T cells stimulated with anti-CD3 (MEM-57) antibody (1 μg/ml). Sample preparation: At the end of stimulation of T cells, perform staining of surface markers (if required) in PBS + 0.1% BSA for 20 min. on ice. Wash with PBS and fix with 2% formaldehyde, 30 min on ice. Wash with PBS and incubate in PBS + 0,1% saponine, 5 min. RT. Incubate the cells in PBS + 1% BSA + 0.03% saponine, 15 min. on ice. Incubate with fluorescence-labeled APA1/1 antibody (1-2 μg/ml) in PBS + 1% BSA + 0.03% saponine in dark, 20 min. RT. Wash with PBS + 1% BSA + 0.03% saponine, resuspend in PBS. Immunocytochemistry: Fixed and permeabilised cells. The antibody can distinguish TCR-stimulated from non-stimulated cells.
• Purity: Purified by protein-A affinity chromatography.
• Conjugation: Unconjugated
• Target: CD3 epsilon (activation epitope)
• Entrez: 916
• UniProt ID: P07766
• Storage: Store at 2-8°C. Do not freeze.
• Buffer/Preservatives: Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
• Alternative names: anti CD 3 antibody, anti Cd247 antibody, anti CD24
• Note: For research use only
• Application notes: Flow cytometry: Intracellular staining; recommended dilution: 1-4 µg/ml; positive control: human T cells stimulated with anti-CD3 (MEM-57) antibody (1 μg/ml). Sample preparation: At the end of stimulation of T cells, perform staining of surface markers (if required) in PBS + 0.1% BSA for 20 min. on ice. Wash with PBS and fix with 2% formaldehyde, 30 min on ice. Wash with PBS and incubate in PBS + 0,1% saponine, 5 min. RT. Incubate the cells in PBS + 1% BSA + 0.03% saponine, 15 min. on ice. Incubate with fluorescence-labeled APA1/1 antibody (1-2 μg/ml) in PBS + 1% BSA + 0.03% saponine in dark, 20 min. RT. Wash with PBS + 1% BSA + 0.03% saponine, resuspend in PBS. Immunocytochemistry: Fixed and permeabilised cells. The antibody can distinguish TCR-stimulated from non-stimulated cells.
• Expiration Date: 12 months from date of receipt.

Separation of Jurkat cells stained anti-human CD3 activation epitope (APA1/1) purified antibody (concentration in sample 5 µg/ml, GAM FITC, red-filled) from Jurkat cells unstained by primary antibody (GAM FITC, black-dashed) in flow cytometry analysis (intracellular staining).