Cofilin 2/CFL2 Antibody (monoclonal, 8C13)

• Catalog Number: orb570307
• Category: Antibodies
• Description: Cofilin 2/CFL2 Antibody (monoclonal, 8C13)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 8C13
• Tested applications: FC, ICC, IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Mouse IgG2b
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Cofilin 2/CFL2 (121-153aa KDAIKKKFTGIKHEWQVNGLDDIKDRSTLGEKL), identical to the related mouse sequence.
• Concentration: 0
• Dilution range: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Immunocytochemistry/Immunofluorescence, 2μg/ml Flow Cytometry, 1-3μg/1x106 cells
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 19 kDa
• UniProt ID: Q9Y281
• 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.
• Alternative names: Cofilin-2; Cofilin, muscle isoform; CFL2
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. 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 500μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). 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 50ug of sample under reducing conditions. Lane 1: human HeLa whole cell lysates Lane 2: human U2OS whole cell lysates Lane 3: human HepG2 whole cell lysates Lane 4: human T-47D whole cell lysates Lane 5: human Raji whole cell lysates Lane 6: human placenta tissue lysates Lane 7: human A549 whole cell 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 mouse anti-Cofilin-2 antigen affinity purified monoclonal antibody (Catalog # orb570307) 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for Cofilin-2 at approximately 19KD. The expected band size for Cofilin-2 is at 19KD.

Western blot analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). 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 50ug of sample under reducing conditions. Lane 1: rat heart tissue lysates Lane 2: rat liver tissue lysates Lane 3: rat kidney tissue lysates Lane 4: rat brain tissue lysates Lane 5: mouse heart tissue lysates Lane 6: mouse liver tissue lysates Lane 7: mouse kidney tissue lysates Lane 8: mouse brain tissue lysates Lane 9: mouse NIH/3T3 whole cell 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 mouse anti-Cofilin-2 antigen affinity purified monoclonal antibody (Catalog # orb570307) 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for Cofilin-2 at approximately 19KD. The expected band size for Cofilin-2 is at 19KD.

IHC analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). Cofilin-2 was detected in paraffin-embedded section of human lung cancer tissues. 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 1μg/ml mouse anti-Cofilin-2 Antibody (orb570307) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IHC analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). Cofilin-2 was detected in paraffin-embedded section of human skeletal muscle tissues. 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 1μg/ml mouse anti-Cofilin-2 Antibody (orb570307) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IHC analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). Cofilin-2 was detected in paraffin-embedded section of mouse skeletal muscle tissues. 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 1μg/ml mouse anti-Cofilin-2 Antibody (orb570307) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IHC analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). Cofilin-2 was detected in paraffin-embedded section of rat skeletal muscle tissues. 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 1μg/ml mouse anti-Cofilin-2 Antibody (orb570307) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IF analysis of Cofilin-2 using anti-Cofilin-2 antibody (orb570307). Cofilin-2 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-Cofilin-2 Antibody (orb570307) overnight at 4°C. DyLight®488 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.

Flow Cytometry analysis of A549 cells using anti-Cofilin-2 antibody (orb570307). Overlay histogram showing A549 cells stained with orb570307 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cofilin-2 Antibody (orb570307, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Flow Cytometry analysis of SiHa cells using anti-Cofilin-2 antibody (orb570307). Overlay histogram showing SiHa cells stained with orb570307 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cofilin-2 Antibody (orb570307, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.