Flotillin 2 Mouse Monoclonal Antibody

SKU: orb1152393

Description

Anti-Flotillin 2 Antibody (monoclonal, 4D8A3). Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Research Area

Protein Biochemistry, Signal Transduction

Images & Validation

• Tested Applications: ICC, IF, IHC, WB
• Dilution Range: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG2b
• Clone No.: 4D8A3
• Immunogen: E.coli-derived human Flotillin 2 recombinant protein (Position: K169-K344). Human Flotillin 2 shares 100% amino acid (aa) sequence identity with both and rat Flotillin 2.
• Target: Flotillin-2
• Molecular Weight: 49 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 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

ECS 1; ECS1; Epidermal surface antigen; ESA; ESA1; FLOT2; flotillin 2; M17S1

IF analysis of FLOT2 using anti-FLOT2 antibody. FLOT2 was detected in an immunocytochemical section of SiHa 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 mouse anti-FLOT2 Antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Mouse 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.

IHC analysis of Flotillin 2 using anti-Flotillin 2 antibody. Flotillin 2 was detected in a paraffin-embedded section of human placenta 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 2 µg/ml mouse anti-Flotillin 2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of Flotillin 2 using anti-Flotillin 2 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 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human A375 whole cell lysates, Lane 4: human A549 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: mouse brain tissue lysates, Lane 7: mouse lung tissue 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 mouse anti-Flotillin 2 antigen affinity purified monoclonal 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-mouse 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 Flotillin 2 at approximately 49 kDa. The expected band size for Flotillin 2 is at 47 kDa.

Quick Database Links

Gene Symbol

Flotillin-2

UniProt

Q14254