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Catalog Number | orb670293 |
---|---|
Category | Antibodies |
Description | C3 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human C3 recombinant protein (Position: Q1277-E1538). Human C3 shares 73.7 % and 76.3% amino acid (aa) sequence identity with mouse and rat C3, respectively. |
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, Rat Flow Cytometry, 1-3μg/1x106 cells, Human Direct ELISA, 0.1-0.5μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 128 kDa, 187 kDa |
UniProt ID | P01024 |
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 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of C3 using anti-C3 antibody (orb670293). 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 30ug of sample under reducing conditions. Lane 1: rat liver tissue lysates, Lane 2: rat plasma lysates, Lane 3: mouse liver tissue lysates, Lane 4: mouse plasma 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-C3 antigen affinity purified polyclonal antibody (Catalog # orb670293) 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 C3 at approximately 128KD/187KD. The expected band size for C3 is at 187KD.
Flow Cytometry analysis of 293T cells using anti-C3 antibody (orb670293). Overlay histogram showing 293T cells stained with orb670293 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-C3 Antibody (orb670293, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
ELISA, IHC-P, WB | |
Human, Mouse, Rat | |
Rabbit | |
Polyclonal | |
Unconjugated |
ELISA, IF, IHC | |
Human | |
Rabbit | |
Polyclonal | |
Unconjugated |
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