Anti-CD3 epsilon/CD3E Antibody

• Catalog Number: orb196263
• Category: Antibodies
• Description: Anti-CD3 epsilon/CD3E Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ICC, IF, IHC, IHC-Fr, WB
• Reactivity: Gallus, Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human CD3 epsilon recombinant protein (Position: D23-I207). Human CD3 epsilon shares 65% amino acid (aa) sequence identity with mouse CD3 epsilon.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 23 kDa
• UniProt ID: P07766
• 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.
• Alternative names: T-cell surface glycoprotein CD3 epsilon chain; T-c
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, Chicken Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Mouse, Rat Immunocytochemistry , 0.5-1μg/ml, Human Immunofluorescence, 2μg/ml, Human, Mouse, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of CD3E and CD20 using anti-CD3E antibody and anti-CD20 antibody. CD3E and CD20 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/mL rabbit anti-CD3E antibody and mouse anti-CD20 antibody overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG, DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of CD3E and CD68 using anti-CD3E antibody and anti-CD68 antibody. CD3E and CD68 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/mL rabbit anti-CD3E Antibody and mouse anti CD68 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG and Biotin conjugated goat anti-rabbit IgG were used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody. CD3 Epsilon was detected in frozen section of mouse spleen tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD3 Epsilon Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody. CD3 Epsilon was detected in frozen section of rat spleen tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD3 Epsilon Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody. CD3 Epsilon was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD3 Epsilon Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody. CD3 Epsilon was detected in paraffin-embedded section of mouse spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD3 Epsilon Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody. CD3 Epsilon was detected in paraffin-embedded section of rat spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD3 Epsilon Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of CD3E using anti-CD3E antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human MOLT-4 whole cell lysates, Lane 2: human Jurkat whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD3E antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CD3E at approximately 23 kDa. The expected band size for CD3E is at 23 kDa.