Anti-EGF Antibody

• Catalog Number: orb402347
• Category: Antibodies
• Description: Anti-EGF Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Rat
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived rat EGF recombinant protein (Position: N974-R1026). Rat EGF shares 69.8% and 77.4% amino acid (aa) sequence identity with human and mouse EGF, respectively.
• 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: 6 kDa
• UniProt ID: P07522
• 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: Pro-epidermal growth factor; EGF; Epidermal growth
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Rat ELISA(Cap) , 1-5μg/ml, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of EGF using anti-EGF antibody. EGF was detected in paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-EGF 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 EGF using anti-EGF antibody. EGF was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-EGF 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.

Sandwich ELISA - Recombinant rat EGF protein standard curve. Use in combination with reagents from Rat EGF ELISA Kit EZ-Set (DIY Antibody Pairs).

Western blot analysis of EGF using anti-EGF antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: recombinant rat EGF protein 1 ng. 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-EGF 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 1:10000 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 EGF at approximately 6KD. The expected band size for EGF is at 6KD.