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CEBP Alpha/CEBPA Rabbit Polyclonal Antibody
Description
Research Area
Images & Validation
−| Tested Applications | ELISA, FC, ICC, IF, WB |
|---|---|
| Dilution Range | Western blot, 0.25-0.5μg/ml, Human Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Human ELISA, 0.1-0.5μg/ml, - |
| Reactivity | Human |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | Rabbit IgG |
| Immunogen | E.coli-derived human CEBP Alpha/CEBPA recombinant protein (Position: Y138-Q221). |
| Target | CCAAT/enhancer-binding protein alpha |
| Molecular Weight | 42 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, 0.2mg Na2HPO4, 0.05mg NaN3. |
| Concentration | 500 µg/ml |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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Flow Cytometry analysis of CACO-2 cells using anti-CEBP Alpha/CEBPA antibody. Overlay histogram showing CACO-2 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CEBP Alpha/CEBPA 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.

IF analysis of CEBP Alpha/CEBPA using anti-CEBP Alpha/CEBPA antibody. CEBP Alpha/CEBPA was detected in immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-CEBP Alpha/CEBPA Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of CEBP using anti-CEBP 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: human THP-1 whole cell lysates, Lane 2: human U-937 whole cell lysates, Lane 3: human HepG2 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-CEBP 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 CEBP at approximately 42 kDa. The expected band size for CEBP is at 37 kDa.
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CEBP Alpha/CEBPA Rabbit Polyclonal Antibody (orb570373)
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