CD33 Rabbit Polyclonal Antibody

SKU: orb371666

Description

CD33 Rabbit Polyclonal Antibody

Research Area

Disease Biomarkers, Immunology & Inflammation, Stem Cell & Developmental Biology

Images & Validation

• Tested Applications: IHC, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
• Reactivity: Human

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived human CD33 recombinant protein (Position: D18-H259). Human CD33 shares 61.6% amino acid (aa) sequence identity with mouse CD33.
• Target: Myeloid cell surface antigen CD33
• Molecular Weight: 60-75 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Myeloid cell surface antigen CD33; Sialic acid-binding Ig-like lectin 3; Siglec-3; gp67; CD33; CD33; SIGLEC3

Flow Cytometry analysis of U937 cells using anti-CD33 antibody.Overlay histogram showing U937 cells (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD33 Antibody (1 µg/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

IHC analysis of CD33 using anti-CD33 antibody. CD33 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 1 µg/ml rabbit anti-CD33 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 CD33 using anti-CD33 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: SKOV3 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-CD33 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 CD33 at approximately 45 kDa. The expected band size for CD33 is at 40 kDa.

Quick Database Links

Gene Symbol

Myeloid cell surface antigen CD33

UniProt

P20138