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Anti-SKAP55/SKAP1 Antibody
Catalog Number: orb334623
| Catalog Number | orb334623 |
|---|---|
| Category | Antibodies |
| Description | Anti-SKAP55/SKAP1 Antibody |
| Species/Host | Rabbit |
| Clonality | Polyclonal |
| Tested applications | IHC, WB |
| Reactivity | Human, Rat |
| Isotype | Rabbit IgG |
| Immunogen | A synthetic peptide corresponding to a sequence in the middle region of human SKAP55, different from the related mouse and rat sequences by two amino acids. |
| Antibody Type | Primary Antibody |
| Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
| Form/Appearance | Lyophilized |
| Conjugation | Unconjugated |
| MW | 60 kDa |
| UniProt ID | Q86WV1 |
| 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. |
| Alternative names | Src kinase-associated phosphoprotein 1; Src family Read more... |
| Note | For research use only |
| Application notes | Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Rat, Human Western blot, 0.1-0.5μg/ml, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml |
| Expiration Date | 12 months from date of receipt. |
IHC analysis of SKAP55 using anti-SKAP55 antibody. SKAP55 was detected in a paraffin-embedded section of rat 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 1 µg/ml rabbit anti-SKAP55 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 SKAP55 using anti-SKAP55 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: PANC whole cell lysates, Lane 2: JURKAT 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-SKAP55 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 SKAP55 at approximately 60 kDa. The expected band size for SKAP55 is at 41 kDa.
