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Antikörper

Phospho-SMAD2 (S465+467) Recombinant Rabbit monoclonal Antibody

Produktgrößen
100μl
£689,00
HA751026-100UL
Über dieses Produkt
SKU:
HA751026
Anwendung:
Western Blot, IHC-Paraffin
Buffer:
PBS (pH7.4).
translate.label.attr.clone:
PSH05-76
Klonalität:
Monoclonal
Konjugat:
Non-conjugated
Host:
Rabbit
Immunogen:
Synthetic phospho-peptide corresponding to residues surrounding Ser465 and 467 of Human SMAD2.
Isotyp:
IgG
Molekulargewicht:
Predicted band size: 52/48 kDa
Physischer Zustand:
Liquid
Reinheit:
Protein A affinity purified.
Versandbedingungen:
Blue Ice
Lagerbedingungen:
Store at +4[o]C after thawing. Aliquot store at -20[o]C. Avoid repeated freeze / thaw cycles.
Hersteller:
Huabio
Typ:
Antibodies: Monoclonal Antibody
Western blot analysis of Phospho-SMAD2 (S465+467) on different lysates with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA751026) at 1/1;000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 starved overnight then treated with 10ng/mL TGF-β1 for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 52/48 kDa Observed band size: 60 kDa Exposure time: 1 minute 20 seconds; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751026) at 1/1;000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HeLa cells treated with or without 20ng/mL TGF-β1 for 15 minutes labeling Phospho-SMAD2 (S465+467) with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA751026) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature; permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature; then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA751026) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488; HA1121) was used as the secondary antibody at 1/1;000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2; red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594; HA1126) was used as the secondary antibody at 1/1;000 dilution.
Immunocytochemistry analysis of NIH/3T3 cells treated with or without 20ng/mL TGF-β1 for 15 minutes labeling Phospho-SMAD2 (S465+467) with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA751026) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature; permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature; then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA751026) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488; HA1121) was used as the secondary antibody at 1/1;000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2; red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594; HA1126) was used as the secondary antibody at 1/1;000 dilution.
Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-Phospho-SMAD2 (S465+467) antibody (HA751026) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature; washed with ddH2O and PBS; and then probed with the primary antibody (HA751026) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.