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CD226 Antibody
Description
Research Area
Images & Validation
−| Tested Applications | ELISA, FC, IHC, WB |
|---|---|
| Dilution range | Western blot, 0.25-0.5μg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Rat ELISA, 0.1-0.5μg/ml, - |
| Reactivity | Mouse, Rat |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | Rabbit IgG |
| Immunogen | E.coli-derived mouse CD226 recombinant protein (Position: E31-L333). |
| Target | CD226 antigen |
| Molecular Weight | 70 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 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4. |
| Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
| Disclaimer | For research use only |
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Flow Cytometry analysis of NRK cells using anti-CD226 antibody. Overlay histogram showing NRK cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD226 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of CD226 using anti-CD226 antibody. CD226 was detected in paraffin-embedded section of mouse lung 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-CD226 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 CD226 using anti-CD226 antibody. CD226 was detected in paraffin-embedded section of rat lung lymphaden 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-CD226 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 CD226 using anti-CD226 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: rat spleen tissue lysates, Lane 2: mouse spleen tissue lysates, Lane 3: mouse thymus tissue lysates, Lane 4: mouse Raw264.7 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-CD226 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 CD226 at approximately 70 KD. The expected band size for CD226 is at 70 KD.
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CD226 Antibody (orb745904)
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