CD5 Antibody (monoclonal, 4E2)

• Catalog Number: orb654261
• Category: Antibodies
• Description: CD5 Antibody (monoclonal, 4E2)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 4E2
• Tested applications: IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG1
• Immunogen: E. coli-derived human CD5 recombinant protein (Position: R25-L495).
• Concentration: 0
• Dilution range: Western blot, 0.1-0.5μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 67 kDa
• UniProt ID: P06127
• Sensitivity: > 5000 cells
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Alternative names: T-cell surface glycoprotein CD5; Lymphocyte antige
• Note: For research use only
• Application notes: Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of CD5 using anti-CD5 antibody (orb654261). 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 50ug of sample under reducing conditions. Lane 1: human Jurkat tissue lysates, Lane 2: human CCRM-CEM whole cell lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-CD5 antigen affinity purified polyclonal antibody (Catalog # orb654261) at 0.5 ?g/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for CD5 at approximately 67KD. The expected band size for CD5 is at 54KD.

IHC analysis of CD5 using anti-CD5 antibody (orb654261). CD5 was detected in paraffin-embedded section of human tonsil cancer 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?g/ml mouse anti-CD5 Antibody (orb654261) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.