bs-0566R [Primary Antibody]
Rabbit  Anti-SMAD7 Rabbit pAb  Polyclonal Antibody
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Host: Rabbit

Target Protein: SMAD7 Rabbit pAb

IR: Immunogen Range:1-100/426

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 4092

Swiss Prot: O15105

Source: KLH conjugated synthetic peptide derived from human Smad7:1-100/426 

Purification: affinity purified by Protein A

Storage: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

Background: The protein encoded by this gene is a nuclear protein that binds the E3 ubiquitin ligase SMURF2. Upon binding, this complex translocates to the cytoplasm, where it interacts with TGF-beta receptor type-1 (TGFBR1), leading to the degradation of both the encoded protein and TGFBR1. Expression of this gene is induced by TGFBR1. Variations in this gene are a cause of susceptibility to colorectal cancer type 3 (CRCS3). Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jun 2010]

Size: 50ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1ug/Test,ICC/IF=1:100

Cross Reactive Species: Human,Mouse,Rat (predicted: Pig,Cow)

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample: Lung(Mouse) Lysate at 30 ug Placenta(Mouse) Lysate at 30 ug Large intestine(Mouse) Lysate at 30 ug Primary: Anti- MADH7/Smad7 (bs-0566R) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 46 kD Observed band size: 50 kD
Sample: Lane 1: Stomach (Mouse) Lysate at 40 ug Lane 2: Spleen (Mouse) Lysate at 40 ug Lane 3: Lung (Mouse) Lysate at 40 ug Primary: Anti-MADH7/Smad7 (bs-0566R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 50 kD Observed band size: 50 kD
Sample: Lane 1: Cerebrum (Mouse) Lysate at 40 ug Lane 2: Cerebrum (Rat) Lysate at 40 ug Lane 3: Stomach (Mouse) Lysate at 40 ug Lane 4: Lung (Mouse) Lysate at 40 ug Lane 5: Kidney (Mouse) Lysate at 40 ug Primary: Anti-MADH7/Smad7 (bs-0566R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 50 kD Observed band size: 50 kD
Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (MADH7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (MADH7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (MADH7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:600 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Tissue/cell: rat kidney 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,C-0005) at 37℃ for 20 min; Incubation: Anti-Smad7/Smad6 Polyclonal Antibody, Unconjugated(bs-0071R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
U-2OS cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (MADH7/Smad7) polyclonal Antibody, Unconjugated (bs-0566R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Blank control: SH-SY5Y. Primary Antibody (green line): Rabbit Anti-MADH7/Smad7 antibody (bs-0566R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : 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-20℃.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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