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Catalog Number | orb692184 |
---|---|
Category | Antibodies |
Description | IL3 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, IHC, WB |
Reactivity | Mouse |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived mouse IL3 recombinant protein (Position: E91-C166). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.25-0.5μg/ml, Mouse Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse Direct ELISA, 0.1-0.5μg/ml, Mouse |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 26 kDa |
UniProt ID | P01586 |
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. |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. 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 IL3 using anti-IL3 antibody (orb692184). 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: mouse spleen tissue 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 rabbit anti-IL3 antigen affinity purified polyclonal antibody (Catalog # orb692184) 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 (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for IL3 at approximately 26KD. The expected band size for IL3 is at 26KD.
IHC analysis of IL3 using anti-IL3 antibody (orb692184). IL3 was detected in paraffin-embedded section of mouse spleen 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 2μg/ml rabbit anti-IL3 Antibody (orb692184) 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) (Catalog # orb90444) with DAB as the chromogen.
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