NFIA Mouse Monoclonal Antibody, Clone: [16H11], Unconjugated

Catalog Number: BYT-ORB527055
Article Name: NFIA Mouse Monoclonal Antibody, Clone: [16H11], Unconjugated
Biozol Catalog Number: BYT-ORB527055
Supplier Catalog Number: orb527055
Alternative Catalog Number: BYT-ORB527055-100
Manufacturer: Biorbyt
Host: Mouse
Category: Antikörper
Application: FC, ICC, IF, IHC, WB
Species Reactivity: Human
Immunogen: A synthetic peptide corresponding to a sequence in the middle region of human NFIA, different from the related mouse sequence by one amino acid, and identical to the related rat sequence.
Conjugation: Unconjugated
Alternative Names: CTF, KIAA1439, NF I/A, NF1 A, NFI A, NFI L, NFIA, Nuclear factor 1 A type, Nuclear factor 1/A, nuclear factor I/A, TGGCA binding protein
Anti-NFIA Antibody (monoclonal, 16H11). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.
Clonality: Monoclonal
Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Clone Designation: [16H11]
Molecular Weight: 62 kDa
UniProt: Q12857
Buffer: Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Form: Lyophilized
Target: Nuclear factor 1 A-type
Application Dilute: Western blot, 0.1-0.5µg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml Immunocytochemistry/Immunofluorescence, 2µg/ml Flow Cytometry (Fixed), 1-3µg/1x10 6 cells
Flow Cytometry analysis of U20S cells using anti-NFIA antibody. Overlay histogram showing U20S 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 mouse anti-NFIA Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-mouse IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was mouse 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.
WB analysis of NFIA using anti-NFIA antibody.Lane 1:human HeLa cell,2
IF analysis of NFIA using anti-NFIA antibody. NFIA was detected in immunocytochemical section of A431 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 2 µg/mL mouse anti-NFIA Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of NFIA using anti-NFIA antibody. NFIA was detected in paraffin-embedded section of human intestinal cancer 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 mouse anti-NFIA Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of NFIA using anti-NFIA antibody. NFIA was detected in paraffin-embedded section of human intestinal cancer 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 mouse anti-NFIA Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of NFIA using anti-NFIA antibody. NFIA was detected in paraffin-embedded section of human tonsil 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 mouse anti-NFIA Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Western blot analysis of NFIA using anti-NFIA 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 50 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human HEK293 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-NFIA antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4C, 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 NFIA at approximately 62KD. The expected