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Smad3 Rabbit pAb
Datasheet   

  • Catalog Number:

    E47R3484

  • Amount:

    100ul

  • Cite ID#:

    EN5553008

  • Price:

    295$

  • Reactivity:

    Human,Mouse,Rat,Pig

  • Predicted Reactivity:

    Chicken,Cow

  • Swiss-Prot No.:

    P84022

  • Form:

    Liquid

  • Altername:

    hMAD 3; hSMAD3; HSPC193; JV15 2; JV152; MAD (mothers against decapentaplegic Drosophila) homolog 3; MAD3; MADH 3; MADH3; Mothers against decapentaplegic homolog 3; Mothers against DPP homolog 3; SMA and MAD related protein 3; SMAD 3; SMAD; SMAD-3; SMAD3_HUMAN.

  • Storage/Stability:

    Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

  • Immunogen:

    KLH conjugated synthetic peptide derived from human Smad3

  • Purification:

    Affinity purified by Protein A

  • Applications:

    WB:1:500-2000,IHC-P:1:100-500,IHC-F:1:100-500,ICC:1:100,IF:1:100-500,Flow-Cyt:1ug/Test,ELISA:1:5000-10000

  • Gene ID:

    4088

  • Subcellular Location:

    Cytoplasm. Nucleus. Note=Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1. Co-localizes with LEMD3 at the nucleus inner membrane. MAPK-mediated phosphorylation appears to have no effect on nuclear import. PDPK1 prevents its nuclear translocation in response to TGF-beta.

  • Antibody type:

    Polyclonal

  • Isotype:

    IgG

  • Immunogen range:

    31-80/425 aa

  • Molecular Weight:

    47 kDa

  • Sample:
    Cerebrum (Mouse) Lysate at 40 ug
    Ovary (Mouse) Lysate at 40 ug
    HT1080 (Human) Cell Lysate at 30 ug
    Jurkat (Human) Cell Lysate at 30 ug
    Primary: Anti- Smad3 at 1/1000 dilution
    Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
    Predicted band size: 52 kD
    Observed band size: 60 kD

  • Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, ) at 37°C for 20 min; Antibody incubation with (Smad3) polyclonal Antibody, Unconjugated 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI was used to stain the cell nuclei.

  • Tissue/cell: mouse lymphoma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
    Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,) at 37℃ for 20 min;
    Incubation: Anti-Smad3 Polyclonal Antibody, Unconjugated 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB() staining

  • Blank control (Black line): HUVEC (Black).
    Primary Antibody (green line): Rabbit Anti-Smad3 antibody
    Dilution: 1μg /10^6 cells;
    Isotype Control Antibody (orange line): Rabbit IgG .
    Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
    Dilution: 1μg /test.
    Protocol
    The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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  • Amount:100ul
  • Price:295.00$

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