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Anti-SMAD5 Antibody EP619Y
Also for SMAD5 (NM_005903)
|A synthetic peptide corresponding to residues near the MH2 domain of human Smad 5 was used as immunogen.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, FC
||IHC-P: Use a concentration of 4 ug/ml; WB: 1:1000; ICC: 1:50; FC: 1:30 - 1:50
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Protein A purified (Protein A or G Sepharose)
|Homo sapiens SMAD family member 5 (SMAD5), transcript variant 1|
|DWFC; JV5-1; MADH5|
|Smad 5 is a member of the Mothers Against Dpp (MAD)-related family of proteins. So far, eight Smads have been identified and can be divided into 3 subgroups based on their structure and function; pathway-restricted, common mediator and inhibitory. Smad 1, 5 and 8 serve as pathway-restricted Smads for the bone morphogenetic proteins (BMPs) signaling pathway (1). Smad 5 is directly activated by BMP type Ia or Ib receptors upon BMP-2 stimulation and transduced BMP-2 signals to the nucleus by forming a complex with Smad 4 (2-3). |
TGF Beta Signaling Pathway
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Western blot - SMAD5 antibody [EP619Y]; Anti-SMAD5 antibody [EP619Y] at 1/1000 dilution + Cos-1 cell lysate at 10 µg.Predicted band size : 52 kDa.Observed band size : 52 kDa.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-SMAD5 antibody [EP619Y](ab40771); ab40771 (4µg/ml) staining SMAD5 in human skin using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclear/cytoplasmic compartments within the stratum granulosum.Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Flow Cytometry-SMAD5 antibody [EP619Y](ab40771); Overlay histogram showing HEK293 cells stained with ab40771 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.