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Anti-NFIA Antibody Picoband™ (monoclonal, 16H11)

     
  •  - Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) ABO14836
    Figure 1. Western blot analysis of NFIA using anti-NFIA antibody (M03531).
    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 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 (Catalog # M03531) 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 # EK1001) with Tanon 5200 system. A specific band was detected for NFIA at approximately 62KD. The expected band size for NFIA is at 62KD.
    detail
  •  - Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) ABO14836
    Figure 2. IHC analysis of NFIA using anti-NFIA antibody (M03531).
    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 (M03531) 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 # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) ABO14836
    Figure 3. IHC analysis of NFIA using anti-NFIA antibody (M03531).
    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 (M03531) 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 # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) ABO14836
    Figure 4. IHC analysis of NFIA using anti-NFIA antibody (M03531).
    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 (M03531) 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 # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) ABO14836
    Figure 5. IF analysis of NFIA using anti-NFIA antibody (M03531).
    NFIA was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-NFIA Antibody (M03531) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.
    detail
  •  - Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) ABO14836
    Figure 6. Flow Cytometry analysis of U20S cells using anti-NFIA antibody (M03531).
    Overlay histogram showing U20S cells stained with M03531 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-NFIA Antibody (M03531, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
    detail
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immuno electron microscopy
  • EIA=Enzyme Immunoassay
WB, IHC, IF, ICC, FC
Primary Accession Q12857
Host Mouse
Isotype Mouse IgG2b
Reactivity Human
Clonality Monoclonal
Format Lyophilized
Description Anti-NFIA Antibody Picoband™ (monoclonal, 16H11) . Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.
Reconstitution Add 0.2ml of distilled water will yield a concentration of 500 µg/ml.
Additional Information
Gene ID 4774
Other Names Nuclear factor 1 A-type, NF1-A, Nuclear factor 1/A, CCAAT-box-binding transcription factor, CTF, Nuclear factor I/A, NF-I/A, NFI-A, TGGCA-binding protein, NFIA, KIAA1439
Calculated MW 62 kDa
Application Details Western blot, 0.1-0.5 µg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1 µg/ml
Immunocytochemistry/Immunofluorescence, 2 µg/ml
Flow Cytometry, 1-3 µg/1x10^6 cells
Subcellular Localization Nucleus
Contents Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clone Names Clone: 16H11
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.
Cross Reactivity No cross-reactivity with other proteins.
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.
Protein Information
Name NFIA
Synonyms KIAA1439
Function Recognizes and binds the palindromic sequence 5'- TTGGCNNNNNGCCAA-3' present in viral and cellular promoters and in the origin of replication of adenovirus type 2. These proteins are individually capable of activating transcription and replication.
Cellular Location Nucleus.
Research Areas
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Background

Nuclear factor 1 A-type is a protein that in humans is encoded by the NFIA gene. Nuclear factor I (NFI) proteins constitute a family of dimericDNA-binding proteins with similar, and possibly identical, DNA-binding specificity. They function as cellular transcription factors and as replication factors for adenovirus DNA replication. Diversity in this protein family is generated by multiple genes, differential splicing, and heterodimerization.

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$ 370.00
Cat# ABO14836
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